[Objective] The aim of this study is to construct eukaryotic expression vector pEGFP-N1-hTERT and observe its expression in eukaryotic cells.[Method]The eukaryotic expression vector pEGFP-N1-hTERT was constructed with...[Objective] The aim of this study is to construct eukaryotic expression vector pEGFP-N1-hTERT and observe its expression in eukaryotic cells.[Method]The eukaryotic expression vector pEGFP-N1-hTERT was constructed with pC1-neo-hTERT and pEGFP-N1 plasmids,and the accuracy of human telomerase reverse transcriptase(hTERT)gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-hTERT into rat fetal neural stem cells(NSCs),the protein localization of human telomerase reverse transcriptase were indirectly observed through green fluorescent protein in the cells,and the correctness of constructed pEGFP-N1-hTERT was certificated by RT-PCR and Western Blot analysis.[Result]The eukaryotic expression vector pEGFP-N1-hTERT had correct structure and could express in eukaryotic cells.[Conclusion]This study laid a foundation for the establishment of immortalized NSCs line in rats.展开更多
[Objective] The aim was to construct the fusion expression vector of polyphosphate kinase（PPK） and green fluorescent protein（GFP） genes.[Method] In this study,the primers were designed based on PPK gene sequence（...[Objective] The aim was to construct the fusion expression vector of polyphosphate kinase（PPK） and green fluorescent protein（GFP） genes.[Method] In this study,the primers were designed based on PPK gene sequence（L03719） of E.coli DH5α in Genbank.Genomic DNA of E.coli DH 5α was extracted as template for the amplification of PPK gene by PCR method.By using In-Fusion@ HD Cloning Kit,the PPK gene was directionally cloned into NcoI site of the pCAMBIA1302 vector.[Result] Sequencing results showed that the 2.0 kb long fragment of PPK gene was inserted into the plant-based expression vector pCAMBIA1302 in front of GFP gene.[Conclusion] The fusion expression vector of PPK and GFP genes were successfully constructed.展开更多
ObjectiveThis study aimed to construct plant expression vector for recombinant human epidermal growth factor （hEGF） and further to provide a basis for the expression of hEGF in peanut hairy root system. MethodAccord...ObjectiveThis study aimed to construct plant expression vector for recombinant human epidermal growth factor （hEGF） and further to provide a basis for the expression of hEGF in peanut hairy root system. MethodAccording to the hEGF sequence in GenBank, hEGF was synthesized artificially; subsequently, hEGF gene was ligated with green fluorescent protein （GFP） gene, and their ligation product was then amplified with primers flanked with corresponding endonuclease cleavage sites, followed by double digestion by Sal I and EcoR I of the amplified products; next, pRI 101 AN DNA was extracted and digested by both Sal I and EcoR I; susequently, the digestion products of hEGF and GFP ligation fragment by Sal I and EcoR I and the digestion products of pRI 101 AN plasmid DNA by Sal I and EcoR I were ligated, and their ligation product was transformed into Escherichia coli XL10-Gold, followed by extraction of DNA from the recombinants exhibiting green fluorescence, which was then identified by enzymatic digestion and PCR, and the verified recombinant plasmid DNA was named pBZG101. ResultHuman epidermal growth factor gene （hEGF） and green fluorescent protein gene （GFP） were successfully ligated, and their ligation fragment was successfully ligated to pRI 101 AN DNA, finally with the acquirement of the plant expression vector for recombinant human epidermal growth factor-（pBZG101）. ConclusionThe plant expression vector for recombinant human epidermal growth factor-（pBZG101）- was successfully constructed in this study.展开更多
[ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein（GFP） with pCIYVV/CP/W, and to develop effective plat virus vector fo...[ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein（GFP） with pCIYVV/CP/W, and to develop effective plat virus vector for plant bioreactor to produce useful protein. [ Method] A section of multiple cloning sites among NIb/CP genes in pCIYVV genome and deoxyribonucleotide polylinker of cleavage recognition sequence containing viral protease Nla were cloned with infectivity full-length cDNA of clover yellow vein virus （CIYVV）, and pCIYVV/CP/W vector was constructed, GFP gene was inserted into pCIyVV/CP/W to construct the pCIYVV/CP/W/GFP vector. The transcription situation of recombinant virus clone was detected by RT-PCR, and targeted gene products expressed by recombinant virus clone were detected with western blot （WB）. [Result] The broad bean seedling inoculated with pCIYVV/CP/W/GFP expressed the same symptom as wild type CIYVV, morbidity was of 100%, the result showed that recombinant virus clone pCIYVV/CP/W/GFP didn＇t suppress, insertion of foreign gene didn＇t destroy the open reading frame of pCIYVV/CP/W. Foreign gene can keep living in F, progeny virus genorne steadily, recombinant virus clone pCIYVV/CP/W/GFP could steadily express GFP in progeny virus at least.[ Conclusion] The useful plant virus vector was provided for useful protein expressing.展开更多
In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was ...In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was constructed and the vector was introduced into tobacco with the agrobacterium-mediated method. PCR results showed that the RrGlu gene was integrated into the tobacco genome.展开更多
[Objectives]This study was conducted to obtain porcine gastrin-releasing peptide(GRP)fusion protein.[Methods]The constructed pET32a(+)-GRP plasmid was transformed into Escherichia coli BL21(DE3)competent cells to obta...[Objectives]This study was conducted to obtain porcine gastrin-releasing peptide(GRP)fusion protein.[Methods]The constructed pET32a(+)-GRP plasmid was transformed into Escherichia coli BL21(DE3)competent cells to obtain the pET32a(+)-GRP-BL21(DE3)fusion protein expression strain,which was induced with 0.5 mM IPTG at 25℃and 150 r/min for 12 h,and the His-tagged GRP fusion protein was detected by SDS-Page gel electrophoresis and Western Blot.[Results]After optimizing the IPTG-induced expression conditions,it was confirmed that the porcine GRP fusion protein was obtained,and the porcine GRP fusion protein was soluble,stable and highly active.[Conclusions]This study lays a foundation for the subsequent preparation of anti-pig GRP antibodies.展开更多
The paper presents the main aspects of contemporary Italian scenario of construction sector,underlying the importance of renoval in the next years,especially concerning energy efficiency retrofit.Moreover,the paper in...The paper presents the main aspects of contemporary Italian scenario of construction sector,underlying the importance of renoval in the next years,especially concerning energy efficiency retrofit.Moreover,the paper includes two important case studies in Italy,housing and schools buildings,which both present great needs of refurbishment.展开更多
Transcription factor NAC102 plays an important role in the abiotic stress responses of plants.In this study,the promoter sequence of 3000 bp located in the upstream of the BjNAC102 gene was cloned from Brassica juncea...Transcription factor NAC102 plays an important role in the abiotic stress responses of plants.In this study,the promoter sequence of 3000 bp located in the upstream of the BjNAC102 gene was cloned from Brassica juncea‘Sichuan Yellow Seed’by using the homologous cloning method.The expression vector of the GUS gene driven by the BjNAC102 promoter was constructed by seamless cloning technology.The results showed that the sequence of the promoter of the BjNAC102 gene contained many cis-acting elements involved in light responsiveness,gibberellinresponsive element,and auxin-responsive element.It was speculated that BjNAC102 played an important role in the abiotic stress response in Brassica juncea.The expression vector of the promoter of the BjNAC102 gene was constructed,which layed a foundation for further studies of the expression pattern of the BjNAC102 gene in Brassica juncea.展开更多
Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells...Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells to synthesized cDNA. The synthesized cDNA encoding pre-miRNA-451 was amplified by polymerase chain reaction （PCR）. The PCR product was separated by electrophoresis on 1% agarose gel and then recovered and purified. The purified cDNA fragments of miRNA-451 precursor sequence was then ligated with vector pLMP for 1 h by using DNA ligase to form pLMP- miRNA-451 plasmid. After that, the pLMP-miRNA-451 plasmid was transformed into E. coli DH5a strain expression system to clone and amplificate. The purified pLMP-miRNA-451 extracted from E. coli DH5a via transformation and clone screening was identificatied with restriction enzyme digestion and DNA sequencing. At last, pLMP-miRNA-451 was transfected into SGC-7901 cells with lip2000. Real-time PCR was used for detection of the miRNA-451, the transfection efficiency was ob- served under fluorescence microscopy and cell counting kit-8 assay was conduced to evaluate the effect of miRNA-451 on SGC-7901 cell proliferation. Results： Our results showed that pLMP-miRNA-451 expression vector was not only constructed successfully and effectively infected SGC-7901 cells, but also could repress the SGC-7901 cell proliferation. Conclusion： The constructed plasmid pLMP-miRNA-451 could used for further studies of miRNA-451 in SGC-7901 cell lines.展开更多
UGPase （UDP-glucose pyrophosphorylase）, one of the primary enzymes concerned with carbohydrate metabolism, catalyzes the formation of UDPG. By inserting the UGPase cDNA fragment cloned from Saccharum officinarum int...UGPase （UDP-glucose pyrophosphorylase）, one of the primary enzymes concerned with carbohydrate metabolism, catalyzes the formation of UDPG. By inserting the UGPase cDNA fragment cloned from Saccharum officinarum into PQE-30, the prokaryotic expression vector of PQE-UGP was successfully constructed. Then the vector plasmid of PQE-UGP was transformed into host bacteria M 15 and the expression of target gene was induced by Isopropyl β-D-1-Thiogalactopyranoside （IPTG）. The research laid foundation for study on the prokaryotic expression of UGPase.展开更多
In China, the trading of construction land quotas has undergone an institutional evolution process characterized by gradual deregulation.In 2021, the Central Committee of the Communist Party of China(CPC) resolved to ...In China, the trading of construction land quotas has undergone an institutional evolution process characterized by gradual deregulation.In 2021, the Central Committee of the Communist Party of China(CPC) resolved to develop a national cross-regional trading mechanism for construction land quotas.Construction land quotas, which have attributes of both public power and private rights, share certain common grounds with the rights of land development, dumping and carbon emission.To build a national trading market for construction land quotas, it is necessary to make clarifications and innovations in macro-level ideas, meso-level mechanisms, and micro-level designs.展开更多
Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by...Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase展开更多
Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant pl...Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant plasmid pMG36e-usp-lacZ.This recombinant plasmid was transformed into both Escherichia coli DH5α and L.lactis MG1363.The enzyme activity,gene sequencing,SDS-PAGE and hereditary stability were assessed and studied.Results The lacZ gene inserted into plasmids pMG36e-usp-lacZ was 99.37% similar to the GenBank sequence,and SDS-PAGE revealed an evident idio-strap at 116 KDa between L.lactis MG1363/pMG36eusp-lacZ in both supernatant and cell samples.β-Galactosidase activity measured 0.225 U/mL in L.lactis pMG36e-usp-lacZ transformants,and its secretion rate was 10%.The plasmid pMG36e-usp-lacZ appeared more stable in MG1363.Conclusion The authors concluded that these new recombinant bacteria well expressed and secreted β-galactosidase,indicating that the β-galactosidase expression system was successfully constructed,and this might provide a new solution for management of lactose intolerance specifically and promote the use of gene-modified organisms as part of the food-grade plasmid in general.展开更多
Connotation of land competitiveness is expatiated from both the narrow sense and broad sense.Evaluation index system of land competitiveness is established according to the 2008 China Statistical Yearbook and 2008 Chi...Connotation of land competitiveness is expatiated from both the narrow sense and broad sense.Evaluation index system of land competitiveness is established according to the 2008 China Statistical Yearbook and 2008 China Land Resources Statistical Yearbook.Efficiency Coefficient Method and Principal Component Analysis Method are used to evaluate the land competitiveness of 31 provincial units in China.Result shows that in the year 2007,land competitiveness gradually decreases from southeast to northwest.The land competitiveness and GDP per unit land have significant negative correlation.The rank of approved new construction land has low positive correlation with the rank of land competitiveness in China.This indicates that there is little correlation between the allocation of regional new construction land and the land use efficiency.Therefore,it is suggested that regional allocation of new construction land should be treated differently based on the evaluation result of land competitiveness.展开更多
In view of the TIN_DDM buffer surface existing in the construction and application of special data type,algorithm efficiency and precision are not matching;the paper applied the rolling ball model in the process of TI...In view of the TIN_DDM buffer surface existing in the construction and application of special data type,algorithm efficiency and precision are not matching;the paper applied the rolling ball model in the process of TIN_DDM buffer surface construction.Based on the precision limitation analysis of rolling ball model,the overall precision control method of rolling ball model has been established.Considering the efficiency requirement of TIN_DDM buffer surface construction,the influence principle of key sampling points and rolling ball radius to TIN_DDM buffer surface construction efficiency has been elaborated,and the rule of identifying key sampling points has also been designed.Afterwards,by erecting the numerical relationship between key sampling points and rolling ball radius,a TIN_DDM buffer surface construction algorithm based on rolling ball acceleration optimization model has been brought forward.The time complexity of the algorithm is O(n).The experiments show that the algorithm could realize the TIN_DDM buffer surface construction with high efficiency,and the algorithm precision is controlled with in 2σ.展开更多
The concept of circular economy has gained recognition as a way to manage waste and conserve resources sustainably,and has the potential to transform the construction industry.This is particularly relevant in the cons...The concept of circular economy has gained recognition as a way to manage waste and conserve resources sustainably,and has the potential to transform the construction industry.This is particularly relevant in the construction industry due to the significant amounts of waste generated during the construction and demolition process.This study examines the perceived importance and effectiveness of strategies related to the circular economy in the construction industry.The data were collected through a survey administered to professionals in the construction sector,capturing their perceptions of various strategies.The results reveal that most strategies received high mean ratings,indicating their perceived significance.Strategies such as waste management and recycling facilities,design for disassembly,and prioritising the use of renewable and sustainable materials were highly valued by the respondents.Additionally,statistical analyses confirmed the significance of these strategies.However,some strategies received comparatively lower ratings,suggesting the need for further attention and improvement.The findings have important implications for policymakers,industry professionals,and stakeholders,guiding decision-making and resource allocation.By prioritising and implementing the identified strategies,stakeholders can drive the adoption of circular economy principles,enhance resource efficiency,and reduce waste in construction practices.Furthermore,this study lays the foundation for future research,highlighting the importance of exploring barriers to implementation,understanding synergies and trade-offs among strategies,conducting longitudinal studies to assess long-term impact,and broadening the participant pool for a more comprehensive understanding.Overall,this study contributes to the growing body of knowledge on the circular economy in the construction industry and provides valuable insights for promoting sustainability and circularity within the sector.展开更多
Long tunnel excavation with tunnel boring ily affected by uncertainties and needs to be adjusted machine （TBM） is a complex and stochastic process. It is eas- according to specific geological conditions in different...Long tunnel excavation with tunnel boring ily affected by uncertainties and needs to be adjusted machine （TBM） is a complex and stochastic process. It is eas- according to specific geological conditions in different tunnel sections, which makes the construction scheduling and management difficult. Based on the rock mass classification, this paper estimates the penetration rate. Using the rate, a cyclic network simulation （CYCLONE） model of TBM boring system is established, and the advance rates under different geological conditions are determined. Then, the impact of different cutter head thrust, which is chosen in reasonable range according to previous experiences, on pro- ject schedule is analyzed. Moreover, the simulation model of mucking system is built to determine the number of muck trains and rail intersections reasonably, regarding the efficiency of muck loading and material transporting. Based on the interaction and interrelation between TBM boring system and mucking system, the combined CY- CLONE model for the entire tunneling process is established. Then reasonable construction schedule, the utilization rate of working resources, and the probability of project completion are obtained through the model programming. At last, a project application shows the feasibility of the presented method.展开更多
The combining site of OKT3 was modeled,and Human Ig LS1 and ND were selected asacceptors of CDRs of OKT3 VL and VH to construct a reshaping antibody against CD3.Bycomparing OKT3,LS1 and ND,with their own family sequen...The combining site of OKT3 was modeled,and Human Ig LS1 and ND were selected asacceptors of CDRs of OKT3 VL and VH to construct a reshaping antibody against CD3.Bycomparing OKT3,LS1 and ND,with their own family sequences,some residues werechanged and the reshaping VL and VH genes were designed.The full VH gene was assem-bled in three steps with eight chemically synthesized oligonucleotide fragments using over-lapping PCR and sequenced.The VH gene was expressed as active protein and inclusion bod-ies in the vectors of pCOMB3 and pGEX-4T-1 by ELISA and Western blot analysis.展开更多
基金Supported by National High-tech Research and Development Program(863Program)of China(2002AA216161)~~
文摘[Objective] The aim of this study is to construct eukaryotic expression vector pEGFP-N1-hTERT and observe its expression in eukaryotic cells.[Method]The eukaryotic expression vector pEGFP-N1-hTERT was constructed with pC1-neo-hTERT and pEGFP-N1 plasmids,and the accuracy of human telomerase reverse transcriptase(hTERT)gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-hTERT into rat fetal neural stem cells(NSCs),the protein localization of human telomerase reverse transcriptase were indirectly observed through green fluorescent protein in the cells,and the correctness of constructed pEGFP-N1-hTERT was certificated by RT-PCR and Western Blot analysis.[Result]The eukaryotic expression vector pEGFP-N1-hTERT had correct structure and could express in eukaryotic cells.[Conclusion]This study laid a foundation for the establishment of immortalized NSCs line in rats.
基金Supported by National Natural Science Foundation of China(31070451)Qianjiang Talent Project of Zhejiang Province(2009R10016)Zhejiang Provincial Natural Science Foundation of China(Y5110067)~~
文摘[Objective] The aim was to construct the fusion expression vector of polyphosphate kinase（PPK） and green fluorescent protein（GFP） genes.[Method] In this study,the primers were designed based on PPK gene sequence（L03719） of E.coli DH5α in Genbank.Genomic DNA of E.coli DH 5α was extracted as template for the amplification of PPK gene by PCR method.By using In-Fusion@ HD Cloning Kit,the PPK gene was directionally cloned into NcoI site of the pCAMBIA1302 vector.[Result] Sequencing results showed that the 2.0 kb long fragment of PPK gene was inserted into the plant-based expression vector pCAMBIA1302 in front of GFP gene.[Conclusion] The fusion expression vector of PPK and GFP genes were successfully constructed.
基金Supported by the Shangdong Natural Science Foundation(ZR2010HQ054)the Ministry of Agriculture Opening Project Fund of Key Laboratory of Rubber Biology/State Key Laboratory Breeding Base of Cultivation&Physiology for Tropical Crops(KLOF1106)the Special Fund for Backbone Teachers and Domestic Visiting Scholars of Shandong Higher Education Institutions9~~
文摘ObjectiveThis study aimed to construct plant expression vector for recombinant human epidermal growth factor （hEGF） and further to provide a basis for the expression of hEGF in peanut hairy root system. MethodAccording to the hEGF sequence in GenBank, hEGF was synthesized artificially; subsequently, hEGF gene was ligated with green fluorescent protein （GFP） gene, and their ligation product was then amplified with primers flanked with corresponding endonuclease cleavage sites, followed by double digestion by Sal I and EcoR I of the amplified products; next, pRI 101 AN DNA was extracted and digested by both Sal I and EcoR I; susequently, the digestion products of hEGF and GFP ligation fragment by Sal I and EcoR I and the digestion products of pRI 101 AN plasmid DNA by Sal I and EcoR I were ligated, and their ligation product was transformed into Escherichia coli XL10-Gold, followed by extraction of DNA from the recombinants exhibiting green fluorescence, which was then identified by enzymatic digestion and PCR, and the verified recombinant plasmid DNA was named pBZG101. ResultHuman epidermal growth factor gene （hEGF） and green fluorescent protein gene （GFP） were successfully ligated, and their ligation fragment was successfully ligated to pRI 101 AN DNA, finally with the acquirement of the plant expression vector for recombinant human epidermal growth factor-（pBZG101）. ConclusionThe plant expression vector for recombinant human epidermal growth factor-（pBZG101）- was successfully constructed in this study.
基金Supported by Natural Science Foundation of Liaoning Province(20072122)Projects Funding of Liaoning Provincial Education Office(05L339)~~
文摘[ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein（GFP） with pCIYVV/CP/W, and to develop effective plat virus vector for plant bioreactor to produce useful protein. [ Method] A section of multiple cloning sites among NIb/CP genes in pCIYVV genome and deoxyribonucleotide polylinker of cleavage recognition sequence containing viral protease Nla were cloned with infectivity full-length cDNA of clover yellow vein virus （CIYVV）, and pCIYVV/CP/W vector was constructed, GFP gene was inserted into pCIyVV/CP/W to construct the pCIYVV/CP/W/GFP vector. The transcription situation of recombinant virus clone was detected by RT-PCR, and targeted gene products expressed by recombinant virus clone were detected with western blot （WB）. [Result] The broad bean seedling inoculated with pCIYVV/CP/W/GFP expressed the same symptom as wild type CIYVV, morbidity was of 100%, the result showed that recombinant virus clone pCIYVV/CP/W/GFP didn＇t suppress, insertion of foreign gene didn＇t destroy the open reading frame of pCIYVV/CP/W. Foreign gene can keep living in F, progeny virus genorne steadily, recombinant virus clone pCIYVV/CP/W/GFP could steadily express GFP in progeny virus at least.[ Conclusion] The useful plant virus vector was provided for useful protein expressing.
文摘In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was constructed and the vector was introduced into tobacco with the agrobacterium-mediated method. PCR results showed that the RrGlu gene was integrated into the tobacco genome.
基金Supported by National Natural Science Foundation of China(31802149)China Postdoctoral Science(2019M651985)+2 种基金Natural Science Foundation of Jiangsu Province(BK20180919)Scientific Research Fund of Nanjing General Hospital of Nanjing Military Command(2016033)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘[Objectives]This study was conducted to obtain porcine gastrin-releasing peptide(GRP)fusion protein.[Methods]The constructed pET32a(+)-GRP plasmid was transformed into Escherichia coli BL21(DE3)competent cells to obtain the pET32a(+)-GRP-BL21(DE3)fusion protein expression strain,which was induced with 0.5 mM IPTG at 25℃and 150 r/min for 12 h,and the His-tagged GRP fusion protein was detected by SDS-Page gel electrophoresis and Western Blot.[Results]After optimizing the IPTG-induced expression conditions,it was confirmed that the porcine GRP fusion protein was obtained,and the porcine GRP fusion protein was soluble,stable and highly active.[Conclusions]This study lays a foundation for the subsequent preparation of anti-pig GRP antibodies.
文摘The paper presents the main aspects of contemporary Italian scenario of construction sector,underlying the importance of renoval in the next years,especially concerning energy efficiency retrofit.Moreover,the paper includes two important case studies in Italy,housing and schools buildings,which both present great needs of refurbishment.
基金Supported by Natural Science Foundation of Hunan Province(2023JJ50083,2023JJ50084)Excellent Youth Project of Hunan Provincial Department of Education(22B0844)Hunan Provincial Graduate Research Innovation Project(CX20231274)。
文摘Transcription factor NAC102 plays an important role in the abiotic stress responses of plants.In this study,the promoter sequence of 3000 bp located in the upstream of the BjNAC102 gene was cloned from Brassica juncea‘Sichuan Yellow Seed’by using the homologous cloning method.The expression vector of the GUS gene driven by the BjNAC102 promoter was constructed by seamless cloning technology.The results showed that the sequence of the promoter of the BjNAC102 gene contained many cis-acting elements involved in light responsiveness,gibberellinresponsive element,and auxin-responsive element.It was speculated that BjNAC102 played an important role in the abiotic stress response in Brassica juncea.The expression vector of the promoter of the BjNAC102 gene was constructed,which layed a foundation for further studies of the expression pattern of the BjNAC102 gene in Brassica juncea.
基金supported by a grant from the Natural Science Foundation of China (No. 8100098)
文摘Objective： The aim of the study was to construct miRNA-451 expression vector pLMP-miRNA-451 which could help identify the functions of miRNA-451 in SGC-7901 cell. Methods： Total RNA was extracted from SGC-7901 cells to synthesized cDNA. The synthesized cDNA encoding pre-miRNA-451 was amplified by polymerase chain reaction （PCR）. The PCR product was separated by electrophoresis on 1% agarose gel and then recovered and purified. The purified cDNA fragments of miRNA-451 precursor sequence was then ligated with vector pLMP for 1 h by using DNA ligase to form pLMP- miRNA-451 plasmid. After that, the pLMP-miRNA-451 plasmid was transformed into E. coli DH5a strain expression system to clone and amplificate. The purified pLMP-miRNA-451 extracted from E. coli DH5a via transformation and clone screening was identificatied with restriction enzyme digestion and DNA sequencing. At last, pLMP-miRNA-451 was transfected into SGC-7901 cells with lip2000. Real-time PCR was used for detection of the miRNA-451, the transfection efficiency was ob- served under fluorescence microscopy and cell counting kit-8 assay was conduced to evaluate the effect of miRNA-451 on SGC-7901 cell proliferation. Results： Our results showed that pLMP-miRNA-451 expression vector was not only constructed successfully and effectively infected SGC-7901 cells, but also could repress the SGC-7901 cell proliferation. Conclusion： The constructed plasmid pLMP-miRNA-451 could used for further studies of miRNA-451 in SGC-7901 cell lines.
文摘UGPase （UDP-glucose pyrophosphorylase）, one of the primary enzymes concerned with carbohydrate metabolism, catalyzes the formation of UDPG. By inserting the UGPase cDNA fragment cloned from Saccharum officinarum into PQE-30, the prokaryotic expression vector of PQE-UGP was successfully constructed. Then the vector plasmid of PQE-UGP was transformed into host bacteria M 15 and the expression of target gene was induced by Isopropyl β-D-1-Thiogalactopyranoside （IPTG）. The research laid foundation for study on the prokaryotic expression of UGPase.
基金a phased research achievement of Sichuan's social science programming project in 2019 titled“Overall Planning of Public Power and Expression of Private Rights in Inter-provincial Trading of Construction Land Quotas of Impoverished Regions”(project number:SC19B093)。
文摘In China, the trading of construction land quotas has undergone an institutional evolution process characterized by gradual deregulation.In 2021, the Central Committee of the Communist Party of China(CPC) resolved to develop a national cross-regional trading mechanism for construction land quotas.Construction land quotas, which have attributes of both public power and private rights, share certain common grounds with the rights of land development, dumping and carbon emission.To build a national trading market for construction land quotas, it is necessary to make clarifications and innovations in macro-level ideas, meso-level mechanisms, and micro-level designs.
文摘Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase
基金supported by the National Science Foundation of China (NO. 30800910)
文摘Objective This study is to examine the secretion effects of β-galactosidase in Lactococcus lactis.Methods The usp45 and β-galactosidase genes were cloned and inserted into plasmid pMG36e to obtain the recombinant plasmid pMG36e-usp-lacZ.This recombinant plasmid was transformed into both Escherichia coli DH5α and L.lactis MG1363.The enzyme activity,gene sequencing,SDS-PAGE and hereditary stability were assessed and studied.Results The lacZ gene inserted into plasmids pMG36e-usp-lacZ was 99.37% similar to the GenBank sequence,and SDS-PAGE revealed an evident idio-strap at 116 KDa between L.lactis MG1363/pMG36eusp-lacZ in both supernatant and cell samples.β-Galactosidase activity measured 0.225 U/mL in L.lactis pMG36e-usp-lacZ transformants,and its secretion rate was 10%.The plasmid pMG36e-usp-lacZ appeared more stable in MG1363.Conclusion The authors concluded that these new recombinant bacteria well expressed and secreted β-galactosidase,indicating that the β-galactosidase expression system was successfully constructed,and this might provide a new solution for management of lactose intolerance specifically and promote the use of gene-modified organisms as part of the food-grade plasmid in general.
基金Supported by the Natural Science Foundation of China (70803032)
文摘Connotation of land competitiveness is expatiated from both the narrow sense and broad sense.Evaluation index system of land competitiveness is established according to the 2008 China Statistical Yearbook and 2008 China Land Resources Statistical Yearbook.Efficiency Coefficient Method and Principal Component Analysis Method are used to evaluate the land competitiveness of 31 provincial units in China.Result shows that in the year 2007,land competitiveness gradually decreases from southeast to northwest.The land competitiveness and GDP per unit land have significant negative correlation.The rank of approved new construction land has low positive correlation with the rank of land competitiveness in China.This indicates that there is little correlation between the allocation of regional new construction land and the land use efficiency.Therefore,it is suggested that regional allocation of new construction land should be treated differently based on the evaluation result of land competitiveness.
基金National Natural Science Foundation of China(Nos.41601498,41471380)National Key R&D Program of China(No.2017YFC1405505)。
文摘In view of the TIN_DDM buffer surface existing in the construction and application of special data type,algorithm efficiency and precision are not matching;the paper applied the rolling ball model in the process of TIN_DDM buffer surface construction.Based on the precision limitation analysis of rolling ball model,the overall precision control method of rolling ball model has been established.Considering the efficiency requirement of TIN_DDM buffer surface construction,the influence principle of key sampling points and rolling ball radius to TIN_DDM buffer surface construction efficiency has been elaborated,and the rule of identifying key sampling points has also been designed.Afterwards,by erecting the numerical relationship between key sampling points and rolling ball radius,a TIN_DDM buffer surface construction algorithm based on rolling ball acceleration optimization model has been brought forward.The time complexity of the algorithm is O(n).The experiments show that the algorithm could realize the TIN_DDM buffer surface construction with high efficiency,and the algorithm precision is controlled with in 2σ.
文摘The concept of circular economy has gained recognition as a way to manage waste and conserve resources sustainably,and has the potential to transform the construction industry.This is particularly relevant in the construction industry due to the significant amounts of waste generated during the construction and demolition process.This study examines the perceived importance and effectiveness of strategies related to the circular economy in the construction industry.The data were collected through a survey administered to professionals in the construction sector,capturing their perceptions of various strategies.The results reveal that most strategies received high mean ratings,indicating their perceived significance.Strategies such as waste management and recycling facilities,design for disassembly,and prioritising the use of renewable and sustainable materials were highly valued by the respondents.Additionally,statistical analyses confirmed the significance of these strategies.However,some strategies received comparatively lower ratings,suggesting the need for further attention and improvement.The findings have important implications for policymakers,industry professionals,and stakeholders,guiding decision-making and resource allocation.By prioritising and implementing the identified strategies,stakeholders can drive the adoption of circular economy principles,enhance resource efficiency,and reduce waste in construction practices.Furthermore,this study lays the foundation for future research,highlighting the importance of exploring barriers to implementation,understanding synergies and trade-offs among strategies,conducting longitudinal studies to assess long-term impact,and broadening the participant pool for a more comprehensive understanding.Overall,this study contributes to the growing body of knowledge on the circular economy in the construction industry and provides valuable insights for promoting sustainability and circularity within the sector.
基金Supported by National Natural Science Foundation of China (No.50709024)Program for New Century Excellent Talents in University (No. NCET-08-0391)
文摘Long tunnel excavation with tunnel boring ily affected by uncertainties and needs to be adjusted machine （TBM） is a complex and stochastic process. It is eas- according to specific geological conditions in different tunnel sections, which makes the construction scheduling and management difficult. Based on the rock mass classification, this paper estimates the penetration rate. Using the rate, a cyclic network simulation （CYCLONE） model of TBM boring system is established, and the advance rates under different geological conditions are determined. Then, the impact of different cutter head thrust, which is chosen in reasonable range according to previous experiences, on pro- ject schedule is analyzed. Moreover, the simulation model of mucking system is built to determine the number of muck trains and rail intersections reasonably, regarding the efficiency of muck loading and material transporting. Based on the interaction and interrelation between TBM boring system and mucking system, the combined CY- CLONE model for the entire tunneling process is established. Then reasonable construction schedule, the utilization rate of working resources, and the probability of project completion are obtained through the model programming. At last, a project application shows the feasibility of the presented method.
基金Supported by National Natural Science Foundation of Chinathe High Technology Research and Development Programme of China.
文摘The combining site of OKT3 was modeled,and Human Ig LS1 and ND were selected asacceptors of CDRs of OKT3 VL and VH to construct a reshaping antibody against CD3.Bycomparing OKT3,LS1 and ND,with their own family sequences,some residues werechanged and the reshaping VL and VH genes were designed.The full VH gene was assem-bled in three steps with eight chemically synthesized oligonucleotide fragments using over-lapping PCR and sequenced.The VH gene was expressed as active protein and inclusion bod-ies in the vectors of pCOMB3 and pGEX-4T-1 by ELISA and Western blot analysis.
