[Objective] To explore the polymorphism of the 3′ flank region of equine IGF-Ⅰ gene. [Method] The 3′ flank region sequences of IGF-Ⅰ gene were amplified from genomic DNA of 270 horses, which included 4 types of Mo...[Objective] To explore the polymorphism of the 3′ flank region of equine IGF-Ⅰ gene. [Method] The 3′ flank region sequences of IGF-Ⅰ gene were amplified from genomic DNA of 270 horses, which included 4 types of Mongolian horse, Sanhe horse and Thoroughbred, and then analyzed by PCR-SSCP. [Result] Three genotypes (AA, BB and AB) were detected by PCR-SSCP and the distribution of genotypes of all research objects except Xinihe horse and Baerhu horse were in line with the "Hardy-Weinberg Law". [Conclusion] There was a polymorphic locus in the 3′ flank region of IGF-Ⅰ gene, which might affect the equine growth and development mechanism. The study is of important theoretical and practical significance to improve the performance and to develop equine industry.展开更多
[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the ...[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the research materials,total DNA was extracted from transgenic Brassica napus by using modified CTAB method.After enzyme digestion and purification,self-joining was made.Two circles of nested PCR and the sequence alignment were carried out.[Result] A fragement with the size of 4.0 kb was amplified ...展开更多
Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, te...Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments （DSAIFF）, was employed to isolate the Mu flanking fragments （MFFs） of miol6. The target site duplications （TSDs） isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and miol6 was mapped in silico onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of miol6 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of miol6 is alternated by Mu insertion.展开更多
The Agrobacterium mediated transgenic rice ( Oryza saliva L.) population with inserts of maize transposon Activator/Dissociation (Ac/Ds) was investigated. DNA sequences flanking the T-DNA were analyzed with inverse PC...The Agrobacterium mediated transgenic rice ( Oryza saliva L.) population with inserts of maize transposon Activator/Dissociation (Ac/Ds) was investigated. DNA sequences flanking the T-DNA were analyzed with inverse PCR. Results showed that 65.4% of the T-DNA was integrated in different locations of rice genome, and some T-DNA flanking sequences were located on certain chromosomes. A number of T-DNA was found to have inserted into protein coding regions. In order to induce transposition of the inserted Ds elements, 354 crosses of Ac x Ds and Ds x Ac were constructed. The excision frequency of Ds element trans-activated by Ac transposase was 22.7% in the F-2 populations, and the transposition was confirmed with analyses of DNA sequences flanking the Ds elements. In addition to the transposition due to 'cut-paste' mechanism, Ds can replicate itself and integrate into a new locus, and inaccurate excisions were also found. A proportion of DNA segments flanking the Ds elements showed no homologies to sequences published in GenBank, of which two were registered under the accession numbers AF355153 and AF355770. The strategy of using transposon tagging for rice genomics study was discussed.展开更多
The genetically modified high-oleic rapeseed （Brassica napus L.） line W-4 was obtained by transforming a binary vector which harbored an inverted repeat expression cassette of fad2 gene into the rapeseed cultivar We...The genetically modified high-oleic rapeseed （Brassica napus L.） line W-4 was obtained by transforming a binary vector which harbored an inverted repeat expression cassette of fad2 gene into the rapeseed cultivar Westar.The transformation was mediated by Agrobacterium.The flanking sequences to both the left and right borders of T-DNA insertion site were amplified by thermal asymmetric interlaced PCR （TAIL-PCR） from the genomic DNA of the transgenic rapeseed line W-4.The flanking sequences to the right border was 290 bp in length and the nucleotide composition was 31.27％ for G＋C content while 68.73％ for A＋T content.The flanking sequence to the left border was 365 bp in length and the G＋C content was 32.6％ and the A＋T content was 67.4％,indicating that the T-DNA was integrated in the A/T-rich region.Further more,sequence alignment analysis showed a deletion of 62 bp including the right border of pCNFIRnos and the integration of the whole left border except a change of G to A.That was to say,the integration of the T-DNA in the transgenic line W-4 not involved in the vector sequences.Based on both flanking sequences as well as the left and right borders of the T-DNA sequences,two pairs of specific primers TLF/TLR and TRF/TRR were designed.Using the primers the event-specific PCR detection method for transgenic rapeseed line W-4 was established.By the PCR,two fragments of 485 and 405 bp were amplified from the W-4 genomic DNA as expected,while no products were amplified from the genomic DNA of other transgenic rapeseed lines and non-transgenic rapeseed line.And by the PCR it is possible to detect the W-4 genomic DNA from a mixed sample of genomic DNA.The limit of the detection for the qualitative PCR assay was 0.1％.The method developed in this work is highly specific,sensitive and suitable for event-specific detection of the transgenic rapeseed line W-4.展开更多
Predicting the cutting forces required for five-axis flank milling is a challenging task due to the difficulties involved in determining the Undeformed Chip Thickness(UCT) and CutterWorkpiece Engagement(CWE). To solve...Predicting the cutting forces required for five-axis flank milling is a challenging task due to the difficulties involved in determining the Undeformed Chip Thickness(UCT) and CutterWorkpiece Engagement(CWE). To solve these problems, this paper presents a new mechanistic cutting force model based on the geometrical analysis of a flank milling process. In the model,the part feature and corresponding cutting location data are taken as input information. The UCT considering cutter runout is calculated according to the instantaneous feed rate of the element cutting edges. A solid-discrete-based method is used to precisely and efficiently identify the CWE between the end mill and the surface being machined. Then, after calibrating the specific force coef-ficients, the mechanistic milling force can be obtained. During the validation process, two practical operations, three-axis flank milling of a vertical surface and five-axis flank milling of a nondevelopable ruled surface, are conducted. Comparisons between predicted and measured cutting forces demonstrate the reliability of the proposed cutting force model.展开更多
In order to ensure machining stability,curvature continuity and smooth cutting force are very important so as to meet the constraints of both cutting force and kinematics of machine tools.For five-axis flank milling,i...In order to ensure machining stability,curvature continuity and smooth cutting force are very important so as to meet the constraints of both cutting force and kinematics of machine tools.For five-axis flank milling,it is difficult to meet both of the constraints because tool path points and tool axis vectors interact with each other.In this paper,multiple relationships between tool path points and tool axis vectors with cutting force and kinematics of machine tools are established,and the strategies of corner-looping milling and clothoidal spirals are combined so as to find feasible solutions under both of the constraints.Tool path parameters are iterated by considering the maximum cutting force and the feasible range of the tool axis vector,and eventually a curvature continuity five-axis flank milling tool path with smooth cutting force is generated.Machining experimental results show that the conditions of cutting force are satisfied,vibration during the process of machining is reduced,and the machining quality of the surface is improved.展开更多
Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studi...Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studies. Herein, we adopted a modified AFLP method to identify and isolate Mu-flanking fragments from maize. The method consists of the following steps： 1） double-digestion of genomic DNA with Bgl II/Msp I and ligation of digested fragments to the Bgl II- and Msp I-adaptors; 2） enrichment of a subset of Bgl II/Msp I fragments followed by selective amplification of the Mu-flanking fragments; 3） simultaneous display of AFLP bands derived from the flanking regions for both insert and native Mu transposons; 4） identification and isolation of AFLP bands resulting from Mu insertions by comparing the banding profiles between Mu-induced mutants and their parental lines; and 5） confirmation of flanking fragments related to these Mu insertions. Using this approach, we have isolated flanking fragment（s） resulting from Mu insertion for every Mu-induced mutant, and one such fragment, M196-FF, is found to contain a partial sequence of the DNA topoisomerase I gene Topl. Moreover, the modified AFLP method including all restriction enzymes, adaptors and primers has been optimized in this study. The modified AFLP method has been proved to be simple and efficient in the isolation of Mu-flanking fragments and will find its usefulness in the functional genomics of maize.展开更多
In five-axis flank milling operations,the intersecting surfaces of different cutting edges create roughness on the milled surfaces that cannot be ignored in situations with strict requirements,especially in aeronautic...In five-axis flank milling operations,the intersecting surfaces of different cutting edges create roughness on the milled surfaces that cannot be ignored in situations with strict requirements,especially in aeronautical manufacturing.To focus on motion problems in milling operations,this paper presents a new model that utilizes elliptical paths as cutting edge trajectories on 3D surface topography machined by peripheral milling.First,the cutter parallel axis offset and location angle are considered,which change the location of the ellipse center and intersection point of the cutting edges.Then,through the proposed model,the predicted surface topography is obtained,and the factors that affect the development tendency of roughness are analyzed.Next,the effects of the cutter location position(CLP)geometric parameters,cutter parallel axis offset and curvature on the roughness are evaluated by a numerical simulation.Finally,machining tests are carried out to validate the model predictions,and the results show that the surface topography predictions correspond well with the experimental results.展开更多
Ruled surfaces found in engineering parts are often blended with a constraint surface,like the blade surface and hub surface of a centrifugal impeller.It is significant to accurately machine these ruled surfaces in fl...Ruled surfaces found in engineering parts are often blended with a constraint surface,like the blade surface and hub surface of a centrifugal impeller.It is significant to accurately machine these ruled surfaces in flank milling with interference-free and fairing tool path,while current models in fulfilling these goals are complex and rare.In this paper,a tool path planning method with optimal cutter locations(CLs)is proposed for 5-axis flank milling of ruled surfaces under multiple geometric constraints.To be specific,a concise three-point contact tool positioning model is firstly developed for a cylindrical cutter.Different tool orientations arise when varying the three contact positions and a tool orientation pool with acceptable cutter-surface deviation is constructed using a meta-heuristic algorithm.Fairing angular curves are derived from candidates in this pool,and then curve registration for cutter tip point on each determined tool axis is performed in respect of interference avoidance and geometric smoothness.On this basis,an adaptive interval determination model is developed for deviation control of interpolated cutter locations.This model is designed to be independent of the CL optimization process so that multiple CLs can be planned simultaneously with parallel computing technique.Finally,tests are performed on representative surfaces and the results show the method has advantages over previous meta-heuristic tool path planning approaches in both machining accuracy and computation time,and receives the best comprehensive performance compared to other multi-constrained methods when machining an impeller.展开更多
This study focuses on the trace and rare earth elements(REE)geochemistry of the Nkporo and Ekenkpon Shales of the Calabar Flank.The main aim is to infer their depositional environment and the degree of their metal enr...This study focuses on the trace and rare earth elements(REE)geochemistry of the Nkporo and Ekenkpon Shales of the Calabar Flank.The main aim is to infer their depositional environment and the degree of their metal enrichment.The shale samples were analyzed using inductively coupled plasma mass spectrometry.The results indicated that the mean concentrations of K,Na,and Fe in Nkporo and Ekenkpon Shales are 1.45,0.4,and 4.17 wt%,and 1.11,0.44,and 5.42 wt%;respectively.The Nkporo Shale is enriched with the following trace elements;P>Mn>Sr>Ba>Zn>Ce>Rb>Zr>V>Cr>Ni and depleted in the following trace elements;Ta>Ge>Sb>Bi>Cd>Ag>Te>In>Hg.While the Ekenkpon Shale is enriched with the following trace elements;P>Mn>Ba>Sr>V>Ce>Zr>Rb>Cr>Zn>Ni and depleted in;Sb>Ge>Bi>Ag>Ce>Te>In>Hg.The concentration of redox-sensitive elements such as V,Ni,Mo,U,Cu,Cr,Re,Cd,Sb,Ti,Mn,and their ratio V/Mo and U/Mo in the black and grey shale samples show different patterns.The REE obtained from the Nkporo and Ekenkpon Shales were PAAS normalized.The Nkporo Shale showed a slightly flat light rare-earth element(LREE),middle rare-earth element(MREE),and heavy rare earth element(HREE)pattern enrichment.Ce/Ce*ranges from 0.95 to 1.09 in Nkporo Shale and 0.67 to 1.40 in Ekenkpon Shale.The Ekenkpon Shale showed a slight LREE,MREE enrichment,and depleted HREE patterns.The Mn contents and U/Mo ratio in Nkporo and Ekenkpon Shales suggests a poor oxygen transitional environment.The V/Mo and V/(V+Ni)ratios indicated that the Nkporo shales were deposited in an anoxic to suboxic conditions and Ekenkpon shales were also deposited under an anoxic to suboxic conditions.The V/Ni ratio indicated that the organic matter in the Nkporo shale is terrigenous while that of the Ekenkpon shales are both terrigenous and marine in origin.展开更多
Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5'\|flanking region. Methods: Genomic sequence from GenBank database (access...Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5'\|flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5'\|flanking region. A 1894 bp fragment of the 5'\|flanking region \{(-1814\} to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to -1344 to +75, -552 to +75 and -337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5\|flanking region\|containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription\|promoting element (promoter) for SNC6 expression resides between 0 to -337, and two transcription\|enhancing elements (enhancer) resides between -337 to -552 and -1344 to -1814, whereas one transcription\|inhibiting element (silencer) exists between -552 to -1344.展开更多
Presents the division of non developable ruled surface into divided small areas and flank milling in these divided areas to improve machining efficiency and machined surface quality by controlling the machining error ...Presents the division of non developable ruled surface into divided small areas and flank milling in these divided areas to improve machining efficiency and machined surface quality by controlling the machining error for each area, and the algorithms developed for generation of tool path and calculation of errors, and concludes from computer simulation results that the algorithms are correct.展开更多
Behavioral studies have shown that flank glands are involved in chemical communication in golden hamsters Mesocricetus auratus but little chemical analysis has been conducted on volatiles arising from these glands. Us...Behavioral studies have shown that flank glands are involved in chemical communication in golden hamsters Mesocricetus auratus but little chemical analysis has been conducted on volatiles arising from these glands. Using gas chromatography-mass spectrometry, we detected compounds from the flank glands of males, only eight of which were also produced in females. Based on these chemical data we performed a number of further experiments. By manipulating light we found that males exposed to short-photoperiods developed smaller flank glands than those exposed to long-photoperiods. Six flank gland volatiles reduced in relative abundance, which possibly coded for reproductive status of males of this seasonally breeding hamster species. Through dyadic encounters, we were able to induce the formation of dominant-subordinate relationships and show that two glandular compounds became high in relative abundance and may function as dominance pheromones. Castration eliminated all male-specific compounds resulting from flank glands, but bilateral ovariectomies only affected one compound in females. Once these ovariectomized females were treated with testosterone, their glandular compounds resembled those of males, suggesting these compounds are under the main control of androgen. Two female putative pheromones, tetradeeanoic acid and hexadecanoic acid, were used in binary choice tests and were both found to attract males over females. Applying a solution of these pheromone compounds to adult males also suppressed their agonistic behavior [Current Zoology 56 （6）： 800-812, 2010].展开更多
TheYinggehaiBasin is aCenozoic high-temperature and high-pressure basinwhere diapir and thermal fluid activitieswere so strong that the deeplysourced natural gas accumulated in the shallowtraps in the diapir structure...TheYinggehaiBasin is aCenozoic high-temperature and high-pressure basinwhere diapir and thermal fluid activitieswere so strong that the deeplysourced natural gas accumulated in the shallowtraps in the diapir structures.Atotal of 11 exploratorywellswere drilled in 8 diapir structures before 2010,but no commercial gas discoveries were made,provoking a hot debate on the possibility of discovering medium-to-large quality gas reservoirs in the middle and deep layers of the diaper structures.A comprehensive analysis of hydrocarbon generation kinetics,reservoir distribution and sealing conditions revealed the following findings.(1)Three gas-charging mechanisms were identified in the study area,namely slow free gas charging and accumulation under buoyancy,slowcharging,evolving and accumulation ofwater soluble gas,and mixed-phase episodic accumulation.(2)In the hightemperature and high-pressure zones in the core of the diapir,the early gas reservoirs experienced multi-stage transformation at later periods,thus the possibility of forming large gas reservoirs dominated by hydrocarbon is small.In the flanks of the diapir,the early gas reservoirsmay bewell preserved at later stages,thus it is possible to discover primary gas pools.In the non-diapir zones,water soluble gas reservoirsmay occur.(3)Three gas accumulation modes may exist in the study area,namely“the mixed-phase transformation mode”of semi-closed overpressure system in the core and periphery of a diapir,“the gas-phase seepage mode”in the flanks of a diapir,and“the water-phase desolvation mode”of a closed over-pressured system in the nondiapir zones.The analysis revealed that the failure of the previous exploratory drillingwas caused mainly by emplacing the wells in the core of the diapir structures.The geochemical behaviors further support that the flanks of a diapir is the favorable place for high-temperature and high-pressure gas accumulation.Exploration practices based on these understandings finally led to the discovery of the large Dongfang 13-1/13-2 gas fields.展开更多
Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. ...Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. The N4081 mutant has a segregation ratio of 1:3.04in average and one T-DNA insertion site according to our assay It was therefore chosen for further analysis. To isolate the joint fragment of T-DNA and plan DNA, the plasmid rescue technique waJs used. pEL-7, one of plasmids from left border of T-DNA, which contained pBR322 was selected from ampicillin plate. The T-DNA fragment of pEL-7 was checked by restriction enzyme analysis and Southern Blot. Restriction analysis confirmed the presence of known sites of EcoRI, PstI and PvuII on it.For confirming the presence of flanking plant DNA in this plasmid, pEL-7 DNA was labeled and hybridized with wild type and mutant plant DNA. The Southern Blot indicated the hybridization band in both of them. Furthermore, the junction of T-DNA/plant DNA was subcloned into bluescript SK+ and sequenced by Applied Biosystem 373A Sequencer. The results showed the 822 bp fragment contained a 274 bp sequence, which is 99.6%homolog (273bp/274 bp) to Ti plasmid pTi 15955 DNA.Ten bp of left 25 bp border repeat were also found in the juction of T-DNA and Plant DNA.Taken together, pEL-7 should contain a joint fragment of T-DNA and flanking plant DNA. This plasmid DNA could be used for the isolation of plant gene, which will be helpful to elucidate the relationship between gene function and plant embryo development.展开更多
Background: Soft song is a low-amplitude song produced by many birds. Recent studies have confirmed that soft song is an aggressive signal. For example, the Brownish-flanked Bush Warblers Cettia fortipes use soft song...Background: Soft song is a low-amplitude song produced by many birds. Recent studies have confirmed that soft song is an aggressive signal. For example, the Brownish-flanked Bush Warblers Cettia fortipes use soft song in male-male conflicts, particularly prior to attacks. Although stable signaling systems require that signals be honest on average,models predict that cheating is an acceptable strategy for some individuals or in some contexts.Methods: This study aimed to test the reliability of soft song as an aggressive signal in the brownish-flanked bush warbler. We used mounted specimens accompanied by broadcast songs or soft songs to simulate a male attempting to invade an existing territory.Results: We found the mounted specimen that coupled playback of soft songs suffered more and quicker attacks from the territory owner and that the relationship between soft song and subsequent attack in the territory owner was far from perfect. We observed territory owners that both over-signaled(i.e., produced soft song but did not attack) and under-signaled(i.e., attacked without producing soft song). Under-signaling territory owners were relatively more commonly than were over-signaling territory owners, particularly in simulated intrusion that coupled playback of soft song with a mount specimen.Conclusions: We discuss the cost of producing soft song and the potential benefit of the unreliable use of soft song and propose a new hypothesis for under-signaling with soft song; i.e., under-signaling territory owners might benefit from taking the initiative in fights.展开更多
A volcano can be defined as a complex system, not least for the hidden clues related to its internal nature. Innovative models grounded in the Artificial Sciences, have been proposed for a novel pattern recognition an...A volcano can be defined as a complex system, not least for the hidden clues related to its internal nature. Innovative models grounded in the Artificial Sciences, have been proposed for a novel pattern recognition analysis at Mt. Etna volcano. The reference monitoring dataset dealt with real data of 28 parameters collected between January 2001 and April 2005, during which the volcano underwent the July-August 2001, October 2002-January 2003 and September 2004-April 2005 flank eruptions. There were 301 eruptive days out of an overall number of 1581 investigated days. The analysis involved successive steps. First, the TWIST algorithm was used to select the most predictive attributes associated with the flank eruption target. During his work, the algorithm TWIST selected 11 characteristics of the input vector: among them SO<sub>2</sub> and CO<sub>2</sub> emissions, and also many other attributes whose linear correlation with the target was very low. A 5 × 2 Cross Validation protocol estimated the sensitivity and specificity of pattern recognition algorithms. Finally, different classification algorithms have been compared to understand if this pattern recognition task may have suitable results and which algorithm performs best. Best results (higher than 97% accuracy) have been obtained after performing advanced Artificial Neural Networks, with a sensitivity and specificity estimates over 97% and 98%, respectively. The present analysis highlights that a suitable monitoring dataset inferred hidden information about volcanic phenomena, whose highly non-linear processes are enhanced.展开更多
文摘[Objective] To explore the polymorphism of the 3′ flank region of equine IGF-Ⅰ gene. [Method] The 3′ flank region sequences of IGF-Ⅰ gene were amplified from genomic DNA of 270 horses, which included 4 types of Mongolian horse, Sanhe horse and Thoroughbred, and then analyzed by PCR-SSCP. [Result] Three genotypes (AA, BB and AB) were detected by PCR-SSCP and the distribution of genotypes of all research objects except Xinihe horse and Baerhu horse were in line with the "Hardy-Weinberg Law". [Conclusion] There was a polymorphic locus in the 3′ flank region of IGF-Ⅰ gene, which might affect the equine growth and development mechanism. The study is of important theoretical and practical significance to improve the performance and to develop equine industry.
基金Supported by National 863 Program of China(2006AA10A113)Natural Science foundation of Zhejiang Province(Y306097)~~
文摘[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the research materials,total DNA was extracted from transgenic Brassica napus by using modified CTAB method.After enzyme digestion and purification,self-joining was made.Two circles of nested PCR and the sequence alignment were carried out.[Result] A fragement with the size of 4.0 kb was amplified ...
基金supported by the High-Tech R&D Program of China(2006AA10A106)the open funds of the National Key Laboratory of Crop Genetic Improvement and China National Fundamental Fund of Personnel Training (J0730649)
文摘Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments （DSAIFF）, was employed to isolate the Mu flanking fragments （MFFs） of miol6. The target site duplications （TSDs） isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and miol6 was mapped in silico onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of miol6 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of miol6 is alternated by Mu insertion.
文摘The Agrobacterium mediated transgenic rice ( Oryza saliva L.) population with inserts of maize transposon Activator/Dissociation (Ac/Ds) was investigated. DNA sequences flanking the T-DNA were analyzed with inverse PCR. Results showed that 65.4% of the T-DNA was integrated in different locations of rice genome, and some T-DNA flanking sequences were located on certain chromosomes. A number of T-DNA was found to have inserted into protein coding regions. In order to induce transposition of the inserted Ds elements, 354 crosses of Ac x Ds and Ds x Ac were constructed. The excision frequency of Ds element trans-activated by Ac transposase was 22.7% in the F-2 populations, and the transposition was confirmed with analyses of DNA sequences flanking the Ds elements. In addition to the transposition due to 'cut-paste' mechanism, Ds can replicate itself and integrate into a new locus, and inaccurate excisions were also found. A proportion of DNA segments flanking the Ds elements showed no homologies to sequences published in GenBank, of which two were registered under the accession numbers AF355153 and AF355770. The strategy of using transposon tagging for rice genomics study was discussed.
基金Supported by Key Agricultural Technology Research and Development Program of Jiangsu Province(BE2009304)Fund for National Rapeseed Research System(CARS13)~~
文摘The genetically modified high-oleic rapeseed （Brassica napus L.） line W-4 was obtained by transforming a binary vector which harbored an inverted repeat expression cassette of fad2 gene into the rapeseed cultivar Westar.The transformation was mediated by Agrobacterium.The flanking sequences to both the left and right borders of T-DNA insertion site were amplified by thermal asymmetric interlaced PCR （TAIL-PCR） from the genomic DNA of the transgenic rapeseed line W-4.The flanking sequences to the right border was 290 bp in length and the nucleotide composition was 31.27％ for G＋C content while 68.73％ for A＋T content.The flanking sequence to the left border was 365 bp in length and the G＋C content was 32.6％ and the A＋T content was 67.4％,indicating that the T-DNA was integrated in the A/T-rich region.Further more,sequence alignment analysis showed a deletion of 62 bp including the right border of pCNFIRnos and the integration of the whole left border except a change of G to A.That was to say,the integration of the T-DNA in the transgenic line W-4 not involved in the vector sequences.Based on both flanking sequences as well as the left and right borders of the T-DNA sequences,two pairs of specific primers TLF/TLR and TRF/TRR were designed.Using the primers the event-specific PCR detection method for transgenic rapeseed line W-4 was established.By the PCR,two fragments of 485 and 405 bp were amplified from the W-4 genomic DNA as expected,while no products were amplified from the genomic DNA of other transgenic rapeseed lines and non-transgenic rapeseed line.And by the PCR it is possible to detect the W-4 genomic DNA from a mixed sample of genomic DNA.The limit of the detection for the qualitative PCR assay was 0.1％.The method developed in this work is highly specific,sensitive and suitable for event-specific detection of the transgenic rapeseed line W-4.
基金co-supported by the Major National S&T Program(No.2014ZX04014-031)the National Natural Science Foundation of China(No.51225503)
文摘Predicting the cutting forces required for five-axis flank milling is a challenging task due to the difficulties involved in determining the Undeformed Chip Thickness(UCT) and CutterWorkpiece Engagement(CWE). To solve these problems, this paper presents a new mechanistic cutting force model based on the geometrical analysis of a flank milling process. In the model,the part feature and corresponding cutting location data are taken as input information. The UCT considering cutter runout is calculated according to the instantaneous feed rate of the element cutting edges. A solid-discrete-based method is used to precisely and efficiently identify the CWE between the end mill and the surface being machined. Then, after calibrating the specific force coef-ficients, the mechanistic milling force can be obtained. During the validation process, two practical operations, three-axis flank milling of a vertical surface and five-axis flank milling of a nondevelopable ruled surface, are conducted. Comparisons between predicted and measured cutting forces demonstrate the reliability of the proposed cutting force model.
基金supported by the National Natural Science Foundation of Chin-China Aerospace Science and Technology Corporation on Advance Manufacturing(No.U1537209)National Natural Science Foundation of China(No.51775278)Jiangsu Province Outstanding Youth Fund of China(No.BK20140036).
文摘In order to ensure machining stability,curvature continuity and smooth cutting force are very important so as to meet the constraints of both cutting force and kinematics of machine tools.For five-axis flank milling,it is difficult to meet both of the constraints because tool path points and tool axis vectors interact with each other.In this paper,multiple relationships between tool path points and tool axis vectors with cutting force and kinematics of machine tools are established,and the strategies of corner-looping milling and clothoidal spirals are combined so as to find feasible solutions under both of the constraints.Tool path parameters are iterated by considering the maximum cutting force and the feasible range of the tool axis vector,and eventually a curvature continuity five-axis flank milling tool path with smooth cutting force is generated.Machining experimental results show that the conditions of cutting force are satisfied,vibration during the process of machining is reduced,and the machining quality of the surface is improved.
文摘Transposable elements have been utilized as mutagens to create mutant libraries for functional genomics. Isolation of genomic segments flanking the insertion Mutator （Mu） is a key step in insertion mutagenesis studies. Herein, we adopted a modified AFLP method to identify and isolate Mu-flanking fragments from maize. The method consists of the following steps： 1） double-digestion of genomic DNA with Bgl II/Msp I and ligation of digested fragments to the Bgl II- and Msp I-adaptors; 2） enrichment of a subset of Bgl II/Msp I fragments followed by selective amplification of the Mu-flanking fragments; 3） simultaneous display of AFLP bands derived from the flanking regions for both insert and native Mu transposons; 4） identification and isolation of AFLP bands resulting from Mu insertions by comparing the banding profiles between Mu-induced mutants and their parental lines; and 5） confirmation of flanking fragments related to these Mu insertions. Using this approach, we have isolated flanking fragment（s） resulting from Mu insertion for every Mu-induced mutant, and one such fragment, M196-FF, is found to contain a partial sequence of the DNA topoisomerase I gene Topl. Moreover, the modified AFLP method including all restriction enzymes, adaptors and primers has been optimized in this study. The modified AFLP method has been proved to be simple and efficient in the isolation of Mu-flanking fragments and will find its usefulness in the functional genomics of maize.
基金financially supported by the Major National S&T Program of China(2017ZX04002001)the Major National S&T Program of China(2016ZX04004004)the National Natural Science Foundation of China(51675301)。
文摘In five-axis flank milling operations,the intersecting surfaces of different cutting edges create roughness on the milled surfaces that cannot be ignored in situations with strict requirements,especially in aeronautical manufacturing.To focus on motion problems in milling operations,this paper presents a new model that utilizes elliptical paths as cutting edge trajectories on 3D surface topography machined by peripheral milling.First,the cutter parallel axis offset and location angle are considered,which change the location of the ellipse center and intersection point of the cutting edges.Then,through the proposed model,the predicted surface topography is obtained,and the factors that affect the development tendency of roughness are analyzed.Next,the effects of the cutter location position(CLP)geometric parameters,cutter parallel axis offset and curvature on the roughness are evaluated by a numerical simulation.Finally,machining tests are carried out to validate the model predictions,and the results show that the surface topography predictions correspond well with the experimental results.
基金supported by the National Natural Science Foundation of China(Nos.U22A20202 and 52205516)the China Postdoctoral Science Foundation(No.2022 M720641)。
文摘Ruled surfaces found in engineering parts are often blended with a constraint surface,like the blade surface and hub surface of a centrifugal impeller.It is significant to accurately machine these ruled surfaces in flank milling with interference-free and fairing tool path,while current models in fulfilling these goals are complex and rare.In this paper,a tool path planning method with optimal cutter locations(CLs)is proposed for 5-axis flank milling of ruled surfaces under multiple geometric constraints.To be specific,a concise three-point contact tool positioning model is firstly developed for a cylindrical cutter.Different tool orientations arise when varying the three contact positions and a tool orientation pool with acceptable cutter-surface deviation is constructed using a meta-heuristic algorithm.Fairing angular curves are derived from candidates in this pool,and then curve registration for cutter tip point on each determined tool axis is performed in respect of interference avoidance and geometric smoothness.On this basis,an adaptive interval determination model is developed for deviation control of interpolated cutter locations.This model is designed to be independent of the CL optimization process so that multiple CLs can be planned simultaneously with parallel computing technique.Finally,tests are performed on representative surfaces and the results show the method has advantages over previous meta-heuristic tool path planning approaches in both machining accuracy and computation time,and receives the best comprehensive performance compared to other multi-constrained methods when machining an impeller.
文摘This study focuses on the trace and rare earth elements(REE)geochemistry of the Nkporo and Ekenkpon Shales of the Calabar Flank.The main aim is to infer their depositional environment and the degree of their metal enrichment.The shale samples were analyzed using inductively coupled plasma mass spectrometry.The results indicated that the mean concentrations of K,Na,and Fe in Nkporo and Ekenkpon Shales are 1.45,0.4,and 4.17 wt%,and 1.11,0.44,and 5.42 wt%;respectively.The Nkporo Shale is enriched with the following trace elements;P>Mn>Sr>Ba>Zn>Ce>Rb>Zr>V>Cr>Ni and depleted in the following trace elements;Ta>Ge>Sb>Bi>Cd>Ag>Te>In>Hg.While the Ekenkpon Shale is enriched with the following trace elements;P>Mn>Ba>Sr>V>Ce>Zr>Rb>Cr>Zn>Ni and depleted in;Sb>Ge>Bi>Ag>Ce>Te>In>Hg.The concentration of redox-sensitive elements such as V,Ni,Mo,U,Cu,Cr,Re,Cd,Sb,Ti,Mn,and their ratio V/Mo and U/Mo in the black and grey shale samples show different patterns.The REE obtained from the Nkporo and Ekenkpon Shales were PAAS normalized.The Nkporo Shale showed a slightly flat light rare-earth element(LREE),middle rare-earth element(MREE),and heavy rare earth element(HREE)pattern enrichment.Ce/Ce*ranges from 0.95 to 1.09 in Nkporo Shale and 0.67 to 1.40 in Ekenkpon Shale.The Ekenkpon Shale showed a slight LREE,MREE enrichment,and depleted HREE patterns.The Mn contents and U/Mo ratio in Nkporo and Ekenkpon Shales suggests a poor oxygen transitional environment.The V/Mo and V/(V+Ni)ratios indicated that the Nkporo shales were deposited in an anoxic to suboxic conditions and Ekenkpon shales were also deposited under an anoxic to suboxic conditions.The V/Ni ratio indicated that the organic matter in the Nkporo shale is terrigenous while that of the Ekenkpon shales are both terrigenous and marine in origin.
文摘Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5'\|flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5'\|flanking region. A 1894 bp fragment of the 5'\|flanking region \{(-1814\} to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to -1344 to +75, -552 to +75 and -337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5\|flanking region\|containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription\|promoting element (promoter) for SNC6 expression resides between 0 to -337, and two transcription\|enhancing elements (enhancer) resides between -337 to -552 and -1344 to -1814, whereas one transcription\|inhibiting element (silencer) exists between -552 to -1344.
文摘Presents the division of non developable ruled surface into divided small areas and flank milling in these divided areas to improve machining efficiency and machined surface quality by controlling the machining error for each area, and the algorithms developed for generation of tool path and calculation of errors, and concludes from computer simulation results that the algorithms are correct.
基金supported by grants from the National Basic Research Program of China (973 Program,no.2010CB833900-2)the Chinese NSF (no.30870297)
文摘Behavioral studies have shown that flank glands are involved in chemical communication in golden hamsters Mesocricetus auratus but little chemical analysis has been conducted on volatiles arising from these glands. Using gas chromatography-mass spectrometry, we detected compounds from the flank glands of males, only eight of which were also produced in females. Based on these chemical data we performed a number of further experiments. By manipulating light we found that males exposed to short-photoperiods developed smaller flank glands than those exposed to long-photoperiods. Six flank gland volatiles reduced in relative abundance, which possibly coded for reproductive status of males of this seasonally breeding hamster species. Through dyadic encounters, we were able to induce the formation of dominant-subordinate relationships and show that two glandular compounds became high in relative abundance and may function as dominance pheromones. Castration eliminated all male-specific compounds resulting from flank glands, but bilateral ovariectomies only affected one compound in females. Once these ovariectomized females were treated with testosterone, their glandular compounds resembled those of males, suggesting these compounds are under the main control of androgen. Two female putative pheromones, tetradeeanoic acid and hexadecanoic acid, were used in binary choice tests and were both found to attract males over females. Applying a solution of these pheromone compounds to adult males also suppressed their agonistic behavior [Current Zoology 56 （6）： 800-812, 2010].
基金National Major Science&Technology Project“Controlling factors for HPHT gas accumulation and exploration orientation in the Ying-Qiong Basin”(No.2011ZX05023-004).
文摘TheYinggehaiBasin is aCenozoic high-temperature and high-pressure basinwhere diapir and thermal fluid activitieswere so strong that the deeplysourced natural gas accumulated in the shallowtraps in the diapir structures.Atotal of 11 exploratorywellswere drilled in 8 diapir structures before 2010,but no commercial gas discoveries were made,provoking a hot debate on the possibility of discovering medium-to-large quality gas reservoirs in the middle and deep layers of the diaper structures.A comprehensive analysis of hydrocarbon generation kinetics,reservoir distribution and sealing conditions revealed the following findings.(1)Three gas-charging mechanisms were identified in the study area,namely slow free gas charging and accumulation under buoyancy,slowcharging,evolving and accumulation ofwater soluble gas,and mixed-phase episodic accumulation.(2)In the hightemperature and high-pressure zones in the core of the diapir,the early gas reservoirs experienced multi-stage transformation at later periods,thus the possibility of forming large gas reservoirs dominated by hydrocarbon is small.In the flanks of the diapir,the early gas reservoirsmay bewell preserved at later stages,thus it is possible to discover primary gas pools.In the non-diapir zones,water soluble gas reservoirsmay occur.(3)Three gas accumulation modes may exist in the study area,namely“the mixed-phase transformation mode”of semi-closed overpressure system in the core and periphery of a diapir,“the gas-phase seepage mode”in the flanks of a diapir,and“the water-phase desolvation mode”of a closed over-pressured system in the nondiapir zones.The analysis revealed that the failure of the previous exploratory drillingwas caused mainly by emplacing the wells in the core of the diapir structures.The geochemical behaviors further support that the flanks of a diapir is the favorable place for high-temperature and high-pressure gas accumulation.Exploration practices based on these understandings finally led to the discovery of the large Dongfang 13-1/13-2 gas fields.
文摘Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. The N4081 mutant has a segregation ratio of 1:3.04in average and one T-DNA insertion site according to our assay It was therefore chosen for further analysis. To isolate the joint fragment of T-DNA and plan DNA, the plasmid rescue technique waJs used. pEL-7, one of plasmids from left border of T-DNA, which contained pBR322 was selected from ampicillin plate. The T-DNA fragment of pEL-7 was checked by restriction enzyme analysis and Southern Blot. Restriction analysis confirmed the presence of known sites of EcoRI, PstI and PvuII on it.For confirming the presence of flanking plant DNA in this plasmid, pEL-7 DNA was labeled and hybridized with wild type and mutant plant DNA. The Southern Blot indicated the hybridization band in both of them. Furthermore, the junction of T-DNA/plant DNA was subcloned into bluescript SK+ and sequenced by Applied Biosystem 373A Sequencer. The results showed the 822 bp fragment contained a 274 bp sequence, which is 99.6%homolog (273bp/274 bp) to Ti plasmid pTi 15955 DNA.Ten bp of left 25 bp border repeat were also found in the juction of T-DNA and Plant DNA.Taken together, pEL-7 should contain a joint fragment of T-DNA and flanking plant DNA. This plasmid DNA could be used for the isolation of plant gene, which will be helpful to elucidate the relationship between gene function and plant embryo development.
基金supported by National Natural Science Foundation of China (No.31172098)the China Postdoctoral Science Foundation (No.2014 M550026)
文摘Background: Soft song is a low-amplitude song produced by many birds. Recent studies have confirmed that soft song is an aggressive signal. For example, the Brownish-flanked Bush Warblers Cettia fortipes use soft song in male-male conflicts, particularly prior to attacks. Although stable signaling systems require that signals be honest on average,models predict that cheating is an acceptable strategy for some individuals or in some contexts.Methods: This study aimed to test the reliability of soft song as an aggressive signal in the brownish-flanked bush warbler. We used mounted specimens accompanied by broadcast songs or soft songs to simulate a male attempting to invade an existing territory.Results: We found the mounted specimen that coupled playback of soft songs suffered more and quicker attacks from the territory owner and that the relationship between soft song and subsequent attack in the territory owner was far from perfect. We observed territory owners that both over-signaled(i.e., produced soft song but did not attack) and under-signaled(i.e., attacked without producing soft song). Under-signaling territory owners were relatively more commonly than were over-signaling territory owners, particularly in simulated intrusion that coupled playback of soft song with a mount specimen.Conclusions: We discuss the cost of producing soft song and the potential benefit of the unreliable use of soft song and propose a new hypothesis for under-signaling with soft song; i.e., under-signaling territory owners might benefit from taking the initiative in fights.
文摘A volcano can be defined as a complex system, not least for the hidden clues related to its internal nature. Innovative models grounded in the Artificial Sciences, have been proposed for a novel pattern recognition analysis at Mt. Etna volcano. The reference monitoring dataset dealt with real data of 28 parameters collected between January 2001 and April 2005, during which the volcano underwent the July-August 2001, October 2002-January 2003 and September 2004-April 2005 flank eruptions. There were 301 eruptive days out of an overall number of 1581 investigated days. The analysis involved successive steps. First, the TWIST algorithm was used to select the most predictive attributes associated with the flank eruption target. During his work, the algorithm TWIST selected 11 characteristics of the input vector: among them SO<sub>2</sub> and CO<sub>2</sub> emissions, and also many other attributes whose linear correlation with the target was very low. A 5 × 2 Cross Validation protocol estimated the sensitivity and specificity of pattern recognition algorithms. Finally, different classification algorithms have been compared to understand if this pattern recognition task may have suitable results and which algorithm performs best. Best results (higher than 97% accuracy) have been obtained after performing advanced Artificial Neural Networks, with a sensitivity and specificity estimates over 97% and 98%, respectively. The present analysis highlights that a suitable monitoring dataset inferred hidden information about volcanic phenomena, whose highly non-linear processes are enhanced.
