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Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97 被引量:113
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作者 Yan Li Zhao-You Tang Sheng-Long Ye Yin-Kun Liu Jie Chen Qiong Xue Jun Chen Dong-Mei Gao Wei-Hua Bao Liver Cancer Institute and Zhongshan Hospital of Fudan University (Former Liver Cancer Institute of Shanghai Medical University),Shanghai 200032,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第5期630-636,共7页
AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, a... AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis. 展开更多
关键词 ALBUMINS Animals Carcinoma Hepatocellular Cell Division Chromosomes clone Cells Flow Cytometry Hepatitis B Hepatitis B Surface Antigens Hepatitis B virus purification Humans Keratin Liver Liver Neoplasms Experimental Male MICE Mice Inbred BALB C Mice Nude Neoplasm Invasiveness Research Support Non-U.S. Gov't Tumor Cells Cultured Virus Integration ALPHA-FETOPROTEINS
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Selection of Media for Hardwood Cuttings Container Seedling-raising of Triploid Clones of Populus tomentosa 被引量:11
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作者 张平冬 王红静 +2 位作者 宋金利 康向阳 朱之悌 《Agricultural Science & Technology》 CAS 2008年第6期104-107,共4页
[Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, r... [Objective] The experiment was aimed to select effective and economical media for container seedling of triploid clones of Populus tomentosa that was carried out. [Method] The sandy loam, peat, perlite, vermiculite, riversand, sludge were taken as media of hardwood cutting and survival rate, seedling height were taken as indexes to select media for container seedling of triploid clones of Populus tomentosa. [Result] Different mixedmedia had great influence on survival rates of container seedlings. Taking peat and vermiculite with the proportion of 5∶2 (M10) or peat ,vermiculite with the proportion of 7∶2 (M11) or sandy loam (M1) as media would generate higher cutting survival rate that was higher than 90.0%. There were significant differece in height increments of container seedlings. Taking sandy loam, peat and vermiculite with the proportion of 6∶2∶2(M5)or sandy loam (M1), seedling height of 60-days the seedling was over 37.0 cm. [Conclusion] According to cost analysis of nursery medium, the optimum medium for hardwood cuttings container seedling-raising of triploid clones of Populus tomentosa was sandy loam. 展开更多
关键词 TRIPLOID cloneS of POPULUS tomentosa HARDWOOD cutting Container seedling-raising Medium
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Diversity of Microflora in Colonic Mucus from Severe Ulcerative Colitis Patients Analyzed by Terminal Restriction Fragment Length Polymorphism and Clone Libraries of Bacterial 16S rRNA Gene Sequences 被引量:1
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作者 I-Nung Huang Yuri Sato +8 位作者 Mitsuo Sakamoto Moriya Ohkuma Shinobu Ohnuma Takeshi Naitoh Chikashi Shibata Akira Horii Junko Nishimura Haruki Kitazawa Tadao Saito 《Advances in Microbiology》 2014年第13期857-870,共14页
Although the gut microflora is thought to be an essential factor in the development of ulcerative colitis (UC), the entire gut microflora occurring in UC remains unknown. Most studies use feces to represent the microf... Although the gut microflora is thought to be an essential factor in the development of ulcerative colitis (UC), the entire gut microflora occurring in UC remains unknown. Most studies use feces to represent the microflora distribution;however, here we analyzed the bacterial diversity in colonic mucus from UC patients receiving colectomy surgery and control patients. The diversity of microflora was investigated using a combination of terminal restriction fragment length polymorphism (T-RFLP) and clone library analyses of the 16S rRNA gene sequences. In the T-RFLP analysis, the number of terminal restriction fragments (T-RFs) decreased significantly in UC patients when compared to control samples. Also in the clone library analysis, the number of operational taxonomic units (OTU) and the Shannon diversity index were reduced significantly in UC patients. These molecular analyses reveal an overall dysbiosis in UC patients. No specific pathogen was found, and a strong negative correlation in relative abundance of bacterial populations was observed between the phyla Bacteroidetes and Firmicutes in the UC patients. This is the first report showing a significant correlation between these two phyla, which may be important characteristics in the pathogenesis of UC. 展开更多
关键词 ULCERATIVE Colitis MICROFLORA Terminal Restriction Fragment Length Polymorphism 16S rRNA Gene clone Library
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Correlations between delayed fluorescence of chlorophyll, metabolism and yield of plants. III. Influence of viral infection on field plants and new technology of clone selection of virus-free planting potato
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作者 Armen B. Avagyan 《Journal of Biophysical Chemistry》 2010年第1期58-63,共6页
At the end of potato plants vegetation virus in-fection induced both decrease in maximum amplitude of delayed fluorescence maximal amplitude and increase half time of its decrease, as well as reduction in the amount o... At the end of potato plants vegetation virus in-fection induced both decrease in maximum amplitude of delayed fluorescence maximal amplitude and increase half time of its decrease, as well as reduction in the amount of stems, plants’ height and assimilation area surface, yield, acceleration of plants development and their early die-off. The differences of DF pa-rameters and yields between strongly and weakly infected plants increase in case of a combined virus infection. In industrial test of the selection of virus-free planting potato by the use of DF parameter, a rise in the yield and de-crease degree of viral infection of crops was obtained. 展开更多
关键词 POTATO Delayed Fluorescence Of CHLOROPHYLL Virus Phonological Stage YIELD clone Selection VIRUS-FREE PLANTING POTATO
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Genetic variation of Norway spruce clones regarding their natural durability, physical and chemical properties
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作者 Ilze Irbe Guna Noldt +3 位作者 Uldis Grinfelds Anrijs Verovkins Aris Jansons Gerald Koch 《Advances in Bioscience and Biotechnology》 2012年第8期1104-1112,共9页
Genetic variation of ten Norway spruce (Picea abies (L.) Karst.) clones regarding their decay resistance against brown rot fungi, as well as physical and chemical properties of clones were investigated. 31- year-old s... Genetic variation of ten Norway spruce (Picea abies (L.) Karst.) clones regarding their decay resistance against brown rot fungi, as well as physical and chemical properties of clones were investigated. 31- year-old spruce clones: 26, 31, A10, A15, A7, B10, B15, B6, V7, V9 were selected across Latvia. The stem diameters of spruce clones varied between 13.0 and 20.9 cm. The wood density of clones ranged from 361 to 443 kg/m3. Klason lignin content, depending on the clone, was between 27.0%-28.9%. Cellular UV microspectrophotometry of the non-infected tracheids displayed the typical lignin distribution with highest absorbance values in the cell corners (abs280 nm 0.80) and compound middle lamellae (abs280 nm 0.48), while secondary wall showed lower lignin absorbance values (abs280 nm0.29 - 0.35). The deposition of phenolic extractives in ray parenchyma and epithelial cells of resin canals were emphasized by a significantly higher UV-absorbance (abs280 nm 0.68 to 0.78) when compared to the cell wall associated lignin. The content of acetone-soluble extractives of spruce clones varied between 1.1% - 1.8%. The x-value (natural durability) for all spruce clones after exposure to C. puteana and P. placentawas >0.90 (durability class 5, not durable). Most of clones after degradation by G. trabeum had x-value > 0.90 with exception of clones B15 and V9 that showed x-value ≤ 0.90 (durability class 4, slightly durable). Natural durability of spruce clones did not correlate with stem diameter, density, content of lignin and extractives. 展开更多
关键词 Norway SPRUCE cloneS Natural Durability Brown ROT Lignin EXTRACTIVES Density UV MICROSPECTROPHOTOMETRY
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Hepatoprotective Effect of Guava (<i>Psidium guajava</i>L.) Leaf Extracts on Ethanol-Induced Injury on Clone 9 Rat Liver Cells 被引量:1
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作者 Hung-Hui Chen Po-Hua Wu +2 位作者 Diana Lo Yun-Chieh Pan Ming-Chang Wu 《Food and Nutrition Sciences》 2011年第9期983-988,共6页
Guava (Psidium guajava L.), a tropical fruit, belongs to Myrtaceae family. Leaves and fruits of guava have been reported to have an anti-diarrheal, hypoglycemic, lipid lowering, anti-bacterial in addition to antioxida... Guava (Psidium guajava L.), a tropical fruit, belongs to Myrtaceae family. Leaves and fruits of guava have been reported to have an anti-diarrheal, hypoglycemic, lipid lowering, anti-bacterial in addition to antioxidant activities. The aim of this study was to investigate several guava leaf extract cytotoxic effects on healthy clone 9 liver cells and its hepatoprotective effects on ethanol-induced heap-toxicity. It was discovered that when the clone 9 liver cells were treated with guava (Psidium guajava Linn.) extracts for 24 hours, there was no retardation of growth as well as when ethanol and acetone extracts at low concentrations 100 μg/mL and 50 μg/mL were administered however cytototoxic effects were detected at higher concentrations. Water and hot water extracts in concentrations lower than or equal to 500 μg/mL revealed no cytotoxic effects. Injury induction to healthy clone 9 liver cells using 5% alcohol concentration for 30 minutes revealed the hepatoprotective properties of guava (Psidium guajava Linn.) extracts. This was significant in concentrations of 100 μg/mL or lower for ethanol and all concentrations for hot water extracts. Hot water extracts showed higher hepatoprotective and lower cytotoxic properties than other extracts. 展开更多
关键词 GUAVA (Psidium Guajava Linn.) Alcohol-Injured CELL Hepatoprotective Properties clone 9 CELL Cytotoxicity
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Bioinformatics and Expression Pattern Analysis of Tomato ns LTP 2-like cDNA full-length Gene Clone
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作者 Zhang Jia He Shan-shan +3 位作者 Zhao Ting-ting Jiang Jing-bin Li Jing-fu Xu Xiang-yang 《Journal of Northeast Agricultural University(English Edition)》 CAS 2019年第1期28-36,共9页
TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulv... TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato. 展开更多
关键词 TOMATO NSLTP 2-like CDNA clone BIOINFORMATICS analysis expression pattern
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淀粉液化芽孢杆菌β-1,3-1,4葡聚糖酶基因的克隆和表达 被引量:9
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作者 吕文平 许梓荣 +2 位作者 李卫芬 孙建义 韩晋辉 《中国兽医学报》 CAS CSCD 北大核心 2005年第3期263-265,共3页
提取淀粉液化芽孢杆菌(Bacillusamyloliquefaciens)基因组,通过PCR克隆了该菌的β-1,3-1,4葡聚糖酶基因全长。结果显示,该基因全长843bp,ORF为717bp,编码239个氨基酸,计算相对分子质量为26800,等电点为6.07。经Blast分析,该序列与基因... 提取淀粉液化芽孢杆菌(Bacillusamyloliquefaciens)基因组,通过PCR克隆了该菌的β-1,3-1,4葡聚糖酶基因全长。结果显示,该基因全长843bp,ORF为717bp,编码239个氨基酸,计算相对分子质量为26800,等电点为6.07。经Blast分析,该序列与基因库中的淀粉液化芽孢杆菌(M15674)同源性最高(97%)。该基因已被GenBank接受(AY205562.1)。用BamH和Xho双酶切目的片段和表达载体pET-30a(+)后相连接,构建了重组表达载体pET-amy,并导入BL21细菌中表达,酶学特性表明SDS-PAGE电泳在26000左右有表达蛋白带,该工程菌总酶活达90.74U/mL,是出发菌的3倍,最适温度在60℃左右,pH值在4~10之间。该工程菌可作为酶活高的理想材料,用以构建耐热性好和酶活高的杂合基因工程菌。 展开更多
关键词 β-1 SDS-PAGE GENBANK PCR Blast pH bp
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牦牛MT-Ⅰ/-Ⅱ cDNA分子克隆及其蛋白质结构分析 被引量:5
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作者 马彬云 任宏伟 +5 位作者 吴建平 徐明旭 张利平 向云 贺鹏飞 蔡欣 《中国生物工程杂志》 CAS CSCD 北大核心 2005年第4期62-68,共7页
利用基因特异引物YMTSP1和YMTSP2,通过RT-PCR从牦牛肝脏组织RNA中克隆出了牦牛 MT-Ⅰ(Genbank Accession No:AY513744)和MT-Ⅱ(Genbank Accession No:AY513745)基因编码区全 长。将牦牛MT-Ⅰ和MT-Ⅱ cDNA序列在CBI上进行同源性搜索发现... 利用基因特异引物YMTSP1和YMTSP2,通过RT-PCR从牦牛肝脏组织RNA中克隆出了牦牛 MT-Ⅰ(Genbank Accession No:AY513744)和MT-Ⅱ(Genbank Accession No:AY513745)基因编码区全 长。将牦牛MT-Ⅰ和MT-Ⅱ cDNA序列在CBI上进行同源性搜索发现,牦牛MT-Ⅰ/-Ⅱ编码区序列在 不同哺乳动物中相当保守。牦牛MT-Ⅰ和MT-Ⅱ编码的MT-Ⅰ和MT-Ⅱ蛋白分别由61个氨基酸组 成,其具有保守的短肽结构如:C-X-C,C-C-X-C-C,C-X-X-C等,其决定MT蛋白分子的整个三维结 构,在分子进化上十分保守。同时对牦牛MT的疏水性和跨膜区分析表明,牦牛MT蛋白可能不 存在跨膜区,也不存在信号肽,是1种非分泌蛋白。并通过同源比较模建,预测和构建了牦牛 MT-Ⅰ和MT-Ⅱ蛋白的分子空间结构,表明牦牛MT-Ⅰ和MT-Ⅱ由α-和β-两个结构域组成,在α-结构 域含有5个Cys短肽结构,β-结构域有4个Cys短肽结构,且2个结构域由保守的三肽序列KKS 相连。 展开更多
关键词 cDNA CDNA Cys MT RNA PCR CBI
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Infectivity Analysis of Cloned Genomic DNA of P1 Agent in vitro 被引量:3
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作者 WEN Li-bin HE Kong-wang YANG Han-chun 《畜牧兽医学报》 CAS CSCD 北大核心 2009年第S1期43-46,共4页
The ultrastructure of porcine kidney(PK)-15 cells was examined after lipofectamine-aided transfection of the molecular clone of the P1 agent.PK-15 cells transfected with the tandem dimer of the P1molecular DNA clone h... The ultrastructure of porcine kidney(PK)-15 cells was examined after lipofectamine-aided transfection of the molecular clone of the P1 agent.PK-15 cells transfected with the tandem dimer of the P1molecular DNA clone had numbers of intracytoplasmic inclusions,and a few cells had intranuclear inclusions.Intracytoplasmic inclusions were round to oval and 0.1-0.3μm in diameter,and intranuclear inclusions,which were more electron dense,were of two general types:the first were round and small(0.1μm approximately)and the second were hexagonal and larger(0.4-0.8μm in diameter).Cells transfected with the tandem dimer of the P1 molecular DNA clone tested positive for P1 DNA at passage 5.This is the first report that the P1 molecular clone has infectivity in vitro and it will provide fundamental materials for further study of the biological characterization of P1. 展开更多
关键词 P1 AGENT molecular clone in vitro infectivity ANALYSIS
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阴道毛滴虫黏附蛋白ap33基因克隆及其表达 被引量:2
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作者 黄慧聪 梁韶晖 +2 位作者 秦茜 潘长旺 谭峰 《寄生虫与医学昆虫学报》 CAS 2005年第1期1-5,共5页
通过提取阴道毛滴虫mRNA ,逆转录合成cDNA ,PCR得到预期长度片段后 ,将其克隆到pUCm-T载体中进行PCR、酶切及测序分析 ,并与GeneBank中核苷酸序列进行同源性分析。再将其克隆至表达载体PET-32a。重组子用酶切、PCR和测序鉴定后 ,转化大... 通过提取阴道毛滴虫mRNA ,逆转录合成cDNA ,PCR得到预期长度片段后 ,将其克隆到pUCm-T载体中进行PCR、酶切及测序分析 ,并与GeneBank中核苷酸序列进行同源性分析。再将其克隆至表达载体PET-32a。重组子用酶切、PCR和测序鉴定后 ,转化大肠杆菌并以异丙基 -B- D- 硫代半乳糖苷 (IPTG)诱导表达。从阴道毛滴虫临床分离株中扩增出 92 7bp的ap33的基因片段 ,构建重组质粒PET- 32a (+) - ap33;IPTG诱导后 ,SDS -PAGE显示表达产物的大小约 5 0kDa。 展开更多
关键词 puCm-T SDS-PAGE PCR mRNA cDNA IPTG
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华支睾吸虫卵黄前体蛋白B基因的识别、序列分析和克隆表达
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作者 黄艳 吴忠道 +2 位作者 吴德 胡旭初 余新炳 《寄生虫与医学昆虫学报》 CAS 2005年第1期20-24,共5页
为识别和克隆华支睾吸虫新基因 ,对华支睾cDNA质粒文库进行随机筛选并测序 ,并利用在线生物信息学工具进行序列分析 ,识别华支睾吸虫未知基因 ,同时根据PGEX -4T- 1多克隆位点及未知基因的序列设计引物 ,PCR扩增目的基因 ,并构建原核重... 为识别和克隆华支睾吸虫新基因 ,对华支睾cDNA质粒文库进行随机筛选并测序 ,并利用在线生物信息学工具进行序列分析 ,识别华支睾吸虫未知基因 ,同时根据PGEX -4T- 1多克隆位点及未知基因的序列设计引物 ,PCR扩增目的基因 ,并构建原核重组质粒。结果发现了CsvpB基因 ,其完整阅读框含 76 2个碱基 ,编码 2 5 4个氨基酸 ,理论分子量为 2 7. 7kDa。序列分析表明 ,CsvpB蛋白与其它物种的卵黄前体蛋白有较高的同源性 ,所构建的重组原核表达质粒PGEX- 4T- 1 vpB经PCR、双酶切及测序证实与目标基因相符。 展开更多
关键词 B PCR cDNA
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烟夜蛾幼虫中肠Bt Cry1Ac毒素受体蛋白cDNA片段的克隆和序列测定 被引量:1
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作者 安世恒 郭线茹 +2 位作者 罗梅浩 蒋金炜 马继盛 《华北农学报》 CSCD 北大核心 2005年第2期84-88,共5页
利用RT-PCR技术扩增烟夜蛾(HelicoverpaassultaGuen啨e)幼虫中肠Bt毒素Cry1Ac受体蛋白APN(N-氨基肽酶,aminopeptidaseN,APN)基因片段,克隆和测序结果表明,测序得到的812bp的片段编码270个氨基酸残基,且该片段在阅读框内。通过同源性分... 利用RT-PCR技术扩增烟夜蛾(HelicoverpaassultaGuen啨e)幼虫中肠Bt毒素Cry1Ac受体蛋白APN(N-氨基肽酶,aminopeptidaseN,APN)基因片段,克隆和测序结果表明,测序得到的812bp的片段编码270个氨基酸残基,且该片段在阅读框内。通过同源性分析发现,其核苷酸序列与棉铃虫(H.armigera)、澳洲棉铃虫(H.punctiger a)、烟芽夜蛾(H.virescens)、舞毒蛾(Lymantriadispar)、小菜蛾(Plutellaxylostella)、印度谷螟(Plodiainterpunctella)RC688品系和HD198品系、烟草天蛾(Manducasexta)和家蚕(Bombyxmori)的Cry1Ac受体蛋白基因的同源性分别为97.0%,90.0%,78.0%,63.5%,55.0%,60.3%,61.2%,55.0%和59.0%。推导的烟夜蛾Cry1Ac受体蛋白基因的氨基酸序列与棉铃虫、烟芽夜蛾、斑实夜蛾、舞毒蛾的氨基酸序列同源性分别为95.6%,81 0%,82 7%和55 7%。该片段编码的氨基酸属于氨肽酶家族,与烟夜蛾对BtCry1Ac毒素的抗性有关。 展开更多
关键词 CDNA PCR inter Bt
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Mongolian Similar Elements Clustering via Immune Clone Algorithm
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作者 Chunhua Chao ZHANG +1 位作者 Yan LIU Wei WU 《Journal of Mathematical Research with Applications》 CSCD 2019年第6期745-754,共10页
Text clustering is an important research issue of clustering technique. It aims to use the similar characteristics or similar expression to group the text so that the texts in the same clusters have the greatest simil... Text clustering is an important research issue of clustering technique. It aims to use the similar characteristics or similar expression to group the text so that the texts in the same clusters have the greatest similarity, and those in different clusters have the greatest dissimilarity.There are many characteristics in Mongolian structure and writing-mode compared with other kinds of characters. By combining K-means and clone immune algorithm, we propose a novel clustering technique called ICKM. Numerical experiments on four elements sets illustrate the validity of our method in the clustering task for Mongolian. 展开更多
关键词 IMMUNE clone algorithm MONGOLIAN similar ELEMENTS CLUSTERING K-MEANS
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河南HIV感染者的HIV-1 B型株tat基因的克隆及序列分析
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作者 孙薏 徐勤枝 +4 位作者 李敬云 隋建丽 李韩平 吴松峰 周平坤 《生物技术通讯》 CAS 2005年第3期245-248,共4页
克隆河南人免疫缺陷病毒(HIV)感染者HIV-1B型株tat基因完整编码框序列,并分析比较其编码产物的序列结构特点。使用重叠PCR技术,从河南省1名HIV-1感染者外周血标本中扩增出tat基因第一和第二外显子并重组为完整的tat基因序列。获得的HIV... 克隆河南人免疫缺陷病毒(HIV)感染者HIV-1B型株tat基因完整编码框序列,并分析比较其编码产物的序列结构特点。使用重叠PCR技术,从河南省1名HIV-1感染者外周血标本中扩增出tat基因第一和第二外显子并重组为完整的tat基因序列。获得的HIV-1B病毒株tat基因,第一外显子为263bp,第二外显子为214bp。将该基因编码产物与其他HIV-1株Tat蛋白经DNA软件编辑并翻译成蛋白质,使用ClustalX1.81进行多序列对比分析发现,第一外显子编码产物的3个保守区域的氨基酸组成大致相同,只有少数氨基酸存在差异。由于Tat蛋白不同病毒株间有高度保守的Cys富集区、核心区和碱性氨基酸富集区,tat基因的克隆为研究其功能并以其为靶点设计和筛选抗艾滋病的药物奠定了基础。 展开更多
关键词 HIV-1 tat B TAT PCR DNA
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3株致病性鸡大肠杆菌P型菌毛结构基因的克隆、序列测定及同源性分析 被引量:1
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作者 苗玉和 王文成 张贵刚 《中国兽医学报》 CAS CSCD 北大核心 2005年第3期250-252,共3页
用鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA做为模板,PCR分别扩增出了0.55kb的P型菌毛结构基因(papA)。将扩增得到的3个papA基因片段分别克隆进pGEM-T○R载体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得... 用鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA做为模板,PCR分别扩增出了0.55kb的P型菌毛结构基因(papA)。将扩增得到的3个papA基因片段分别克隆进pGEM-T○R载体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到含阳性重组子的菌株,提取质粒后用BamH和Sal双酶切进行鉴定。所得阳性重组子进行DNA序列测定,测序结果经DNAStar核酸分析软件包分析比较,结果表明,所构建的克隆质粒中均含有相应完整papA基因,其开放式阅读框架大小为549bp,编码182个氨基酸。此3株菌的P型菌毛结构基因的同源性为98.9%~100%,其中O1株和O78株的P型菌毛基因的ORF序列100%相同,O2株有2个碱基与前两者不同。 展开更多
关键词 P DNA DNA IPTG Star PCR Amp Sal
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黑杨派无性系生长与材性联合选择 被引量:19
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作者 李善文 姜岳忠 +2 位作者 王桂岩 王卫东 乔玉玲 《林业科学》 EI CAS CSCD 北大核心 2005年第2期53-58,共6页
利用 9年生黑杨派 10个无性系试验林材料进行了生长与材性等 14个性状的遗传分析及选择研究。方差分析、遗传参数估测表明 ,无性系间 14个性状差异均达极显著水平 ,各性状无性系重复力在 0 85 2~ 0 987。说明这些无性系在多个性状上... 利用 9年生黑杨派 10个无性系试验林材料进行了生长与材性等 14个性状的遗传分析及选择研究。方差分析、遗传参数估测表明 ,无性系间 14个性状差异均达极显著水平 ,各性状无性系重复力在 0 85 2~ 0 987。说明这些无性系在多个性状上存在广泛遗传差异 ,并且这种差异受较强的遗传控制 ,从中进行多性状遗传改良是可行的。经多性状主成分分析和聚类分析 ,选出卡帕茨、5 0杨、I 6 9三个综合性状表现优良的无性系 ,其中卡帕茨为新选无性系 ,在该试验林中生长量最高 ,材积超I 2 14杨 6 9 7%、超中林 4 6杨 7 9% ,且材质优良 。 展开更多
关键词 46 I-69 广
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Code Clone Detection Method Based on the Combination of Tree-Based and Token-Based Methods
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作者 Ryota Ami Hirohide Haga 《Journal of Software Engineering and Applications》 2017年第13期891-906,共16页
This article proposes the high-speed and high-accuracy code clone detection method based on the combination of tree-based and token-based methods. Existence of duplicated program codes, called code clone, is one of th... This article proposes the high-speed and high-accuracy code clone detection method based on the combination of tree-based and token-based methods. Existence of duplicated program codes, called code clone, is one of the main factors that reduces the quality and maintainability of software. If one code fragment contains faults (bugs) and they are copied and modified to other locations, it is necessary to correct all of them. But it is not easy to find all code clones in large and complex software. Much research efforts have been done for code clone detection. There are mainly two methods for code clone detection. One is token-based and the other is tree-based method. Token-based method is fast and requires less resources. However it cannot detect all kinds of code clones. Tree-based method can detect all kinds of code clones, but it is slow and requires much computing resources. In this paper combination of these two methods was proposed to improve the efficiency and accuracy of detecting code clones. Firstly some candidates of code clones will be extracted by token-based method that is fast and lightweight. Then selected candidates will be checked more precisely by using tree-based method that can find all kinds of code clones. The prototype system was developed. This system accepts source code and tokenizes it in the first step. Then token-based method is applied to this token sequence to find candidates of code clones. After extracting several candidates, selected source codes will be converted into abstract syntax tree (AST) for applying tree-based method. Some sample source codes were used to evaluate the proposed method. This evaluation proved the improvement of efficiency and precision of code clones detecting. 展开更多
关键词 Code clone Token-Based DETECTION Tree-Based DETECTION TREE EDIT Distance
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Genetic Control of Air-Dried Wood Density, Mechanical Properties and Its Implication for Veneer Timber Breeding of New Triploid Clones in Populus tomentosa Carr
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作者 邢新婷 张志毅 《Forestry Studies in China》 CAS 2002年第2期52-60,共9页
The wood samples of 9 triploid clones of Populus tomentosa Carr. taken from a 9 year old clonal test site were analyzed in order to investigate the genetic variation of wood properties, including air dried wood... The wood samples of 9 triploid clones of Populus tomentosa Carr. taken from a 9 year old clonal test site were analyzed in order to investigate the genetic variation of wood properties, including air dried wood density and some mechanical properties. The results showed that significant or extremely significant difference in air dried wood density and the mechanical properties existed among the clones, this means these wood properties were under moderate or strong genetic controls and could be improved by genetic manipulations. The radial and vertical variation patterns of air dried wood density were also studied and the results were found to coordinate with other previous research results. The vertical variation patterns of most mechanical properties within the individual tree also conformed to the general wood theories except the modulus of elasticity and cross section hardness. Among the mechanical properties, modulus of elasticity (MOE) and tangent section hardness were under strong genetic control, with the clonal repeatabilities being 0 90 and 0 80, respectively. However, the clonal repeatabilities of other mechanical properties under study were a little lower than above two indexes. Genetic correlation analysis indicated that super clonal selection and breeding for veneer timber could be realized through indirect selection of wood density and form indexes. 展开更多
关键词 Populus tomentosa Carr. triploid clones air dried wood density mechanical properties genetic variation
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Qualitative analysis of catechins from green tea GMB-4 clone using HPLC and LC-MS/MS 被引量:3
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作者 Erna Susanti Ciptati +2 位作者 Retty Ratnawati Aulanni'am Achmad Rudijanto 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2015年第12期1046-1050,共5页
Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the... Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the column with 10%ethanol.Subsequently,the column was eluted with95%ethanol and evaporated separately.Green tea extract was identified by thin layer chromatography.Catechins were separated by the stationary phase in column chromatography using polyamide with 10%ethanol eluent and 95%ethanol.The results of isolations were analyzed by high performance liquid chromatographic(HPLC)and LCMS/MS.Analysis of catechins by HPLC was done by external standard.Results:Fraction from 10%ethanol showed that four major peaks at retention time of1.663,2.367,2.950 and 4.890,indicated the presence of four catechins components including catechin,epicatechins,gallocatechin and epigallocatechin.Whereas,fraction from 95%ethanol showed two main peaks at retention time of 5.167 and 9.82,which indicated the presence of epigallocatechin gallate(EGCG)and epicatechin gallate(ECG).EGCG(m/z 459),epigallocatechin(m/z 307),ECG(m/z 443),and epicatechin(m/z 291)were isolated and separated successfully using HPLC and LC-MS/MS.Conclusions:The HPLC and LC-MS/MS methods were successfully tuned for the qualitative analysis of green tea extract with EGCG and ECG.Four major catechins were separated and identified by LC-MS/MS,such as EGCG,epigallocatechin,ECG and epicatechin.The result of HPLC analysis showed that EGCG and ECG were main components from catechins isolation of green tea GMB-4 clone. 展开更多
关键词 CATECHINS Green TEA GMB-4 clone LC-MS/MS HPLC
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