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Genomic signatures of selection,local adaptation and production type characterisation of East Adriatic sheep breeds
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作者 Boris Lukic Ino Curik +4 位作者 Ivana Drzaic Vlatko Galić Mario Shihabi LubošVostry Vlatka Cubric-Curik 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2024年第2期546-562,共17页
Background The importance of sheep breeding in the Mediterranean part of the eastern Adriatic has a long tradition since its arrival during the Neolithic migrations.Sheep production system is extensive and generally c... Background The importance of sheep breeding in the Mediterranean part of the eastern Adriatic has a long tradition since its arrival during the Neolithic migrations.Sheep production system is extensive and generally carried out in traditional systems without intensive systematic breeding programmes for high uniform trait production(carcass,wool and milk yield).Therefore,eight indigenous Croatian sheep breeds from eastern Adriatic treated here as metapopulation(EAS),are generally considered as multipurpose breeds(milk,meat and wool),not specialised for a particular type of production,but known for their robustness and resistance to certain environmental conditions.Our objective was to identify genomic regions and genes that exhibit patterns of positive selection signatures,decipher their biological and productive functionality,and provide a"genomic"characterization of EAS adaptation and determine its production type.Results We identified positive selection signatures in EAS using several methods based on reduced local variation,linkage disequilibrium and site frequency spectrum(eROHi,iHS,nSL and CLR).Our analyses identified numerous genomic regions and genes(e.g.,desmosomal cadherin and desmoglein gene families)associated with environmental adaptation and economically important traits.Most candidate genes were related to meat/production and health/immune response traits,while some of the candidate genes discovered were important for domestication and evolutionary processes(e.g.,HOXa gene family and FSIP2).These results were also confirmed by GO and QTL enrichment analysis.Conclusions Our results contribute to a better understanding of the unique adaptive genetic architecture of EAS and define its productive type,ultimately providing a new opportunity for future breeding programmes.At the same time,the numerous genes identified will improve our understanding of ruminant(sheep)robustness and resistance in the harsh and specific Mediterranean environment. 展开更多
关键词 Composite-likelihood ratio East Adriatic sheep Extreme ROH islands genomic selection signatures Integrated haplotype score Number of segregating sites by length
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Isolation and Identification of a Specific cDNA Mapping to the Bam HI-I2 and -LFragments within the Inverted Repeats ofUnique Long Re-gion (IRL) in the Genom e ofMarek′s Disease Herpesvirus (MDV) Oncogenic Strain Beijing-1 被引量:13
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作者 Lu Chun, Wu Jianping, Zhang Xunhai, et al. Lu Chun, Wu Jianping, Zhang Xunhai, et al. Department of Microbiology and Immunology, Nanjing Medical University, Nanjing\ 210029 Acta Universitatis Medicinalis Nanjing, 19(6):447 《The Journal of Biomedical Research》 CAS 1999年第2期73-73,共1页
Objective\ To understand the transcription of BamHI L DNA fragment from genome of strong virulent GA strain of Marek′s disease herpesvirus (MDV) in lymphoblastoid tumor tissue induced by oncogenic strain Beijing 1 ... Objective\ To understand the transcription of BamHI L DNA fragment from genome of strong virulent GA strain of Marek′s disease herpesvirus (MDV) in lymphoblastoid tumor tissue induced by oncogenic strain Beijing 1 (a specific local strain in China) of MDV. Methods\ Two oligonucleotide primers were synthesized according to the reported sequence of \%meq\% gene an ideal oncogenic candidate and our previously determined sequence of BamHI L fragment of Marek′s disease herpesvirus (MDV), respectively. Reverse transcriptase PCR(RT PCR) assay was performed by using these primers and the mRNA as a template which was isolated from visceral lymphoblastoid tumors obtained from chickens artificially infected with strain Beijing 1 of oncogenic MDV. Southern blot molecular hybridization was further carried out to detect the product of RT PCR with digoxigenin labeled nucleotide probe from BamHI I2 and L fragment in the gene library of MDV strain GA, respectively. Results\ Two probes could simultaneously hybridize this cDNA amplified by RT PCR with a length of about 730 bp. Conclusion\ It is suggested that \%meq\% transcription could extend from the right hand end of BamHI I2 to the adjacent BamHI L, and the BamHI L region was likely to be transcribed in MDV induced lymphoblastoid tumors. 展开更多
关键词 cdna IRL in the genom e ofMarek Isolation and Identification of a Specific cdna Mapping to the Bam HI-I2 and LFragments within the Inverted Repeats ofUnique Long Re-gion Oncogenic Strain Beijing-1 s Disease Herpesvirus LONG RE MDV
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Cloning and Sequencing of a Full-Length cDNA Encoding the RuBPCase Small Subunit (RbcS) in Tea (Camellia sinensis) 被引量:3
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作者 YE Ai-hua JIANG Chang-jun +4 位作者 ZHU Lin YU Mei WANG Zhao-xia DENG Wei-wei WEI Chao-lin 《Agricultural Sciences in China》 CAS CSCD 2009年第2期161-166,共6页
This study was aimed to isolate ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) from tea plant [Camellia sinensis (L.) O. Kuntze]. In the study of transcriptional profiling of gene expression ... This study was aimed to isolate ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) from tea plant [Camellia sinensis (L.) O. Kuntze]. In the study of transcriptional profiling of gene expression from tea flower bud development stage by cDNA-AFLP (cDNA amplified fragment length polymorphism), we have isolated some transcript-derived fragments (TDFs) occurring in both the young and mature flower bud. One of them showed a high degree of similarity to RbcS. Based on the fragment, the full length of RbcS with 769-bp (EF011075) cDNA was obtained via rapid amplification of cDNA ends (RACE). It contained an open reading frame of 176 amino acids consisting of a chloroplast transit peptide with 52 amino acids and a mature protein of 124 amino acids. The amino acids sequence presented a high identity to those of other plant RbcS genes. It also contains three conserved domains and a protein kinase C phosphorylation site, one tyrosine kinase phosphorylation site and two N-myristoylation sites. Analysis by RT-PCR showed that the expression of RbcS in tea from high to low was leaf, young stem, young flower bud and mature flower bud, respectively. The isolation of the tea Rubisco small subunit gene establishes a good foundation for further study on the photosynthesis of tea plant. 展开更多
关键词 RBCS TEA full-length cdna
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Current Status and Progresses in Chinese Cotton Genomic Research 被引量:1
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作者 Yu-xian ZHU, Sheng-jian JI, Ying-chun LU, Gang WEI, Jun LI(National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China) 《棉花学报》 CSCD 北大核心 2002年第S1期36-36,共1页
Cotton fiber,a single-celled trichome,30-40mmin length and 15m in cell-wall-thickness,is adifferentiated epidermal cell originated from theouter integument of the ovule.There are severalgroups in China that work on co... Cotton fiber,a single-celled trichome,30-40mmin length and 15m in cell-wall-thickness,is adifferentiated epidermal cell originated from theouter integument of the ovule.There are severalgroups in China that work on cotton genecloning or fiber improvement usingblotechnological approaches.Dr.Xiaoya 展开更多
关键词 COTTON COTTON ELONGATION length genomic originated MUTANT SILKWORM preferentially UPLAND
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Construction of a Normalized Full-Length cDNA Library of Sesame Developing Seed by DSN and SMART^(TM) 被引量:8
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作者 KE Tao DONG Cai-hua +3 位作者 MAO Han ZHAO Ying-zhong LIU Hong-yan LIU Sheng-yi 《Agricultural Sciences in China》 CAS CSCD 2011年第7期1004-1009,共6页
Sesame (Sesamue indicum L.) is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultiva... Sesame (Sesamue indicum L.) is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultivar of sesame Zhongzhi 14, during its oil accumulation stages. It combined switching mechanism at 5?end of RNA transcript (SMART) technique and duplex-specific nuclease (DSN) normalization methods. Double-stranded cDNAs were synthesized from mRNAs, processed by normalization and Sfi I restriction endonuclease, and finally the cDNAs were ligated to pDNR-LIB vector. The ligation mixture was transformed into Escherichia coli DH10B by electroporation. The capacity of the library was 1.0?06 clones in this library. Gel electrophoresis results indicated the fragments ranged from 700 to 2 000 bp, with the average size of 1 800 bp. Random picking clones showed that the recombination rate was 100%. The results showed that the cDNA library constructed successfully was a full-length library with high quality, and could be used to screen the genes related to development of oil synthesis. 展开更多
关键词 DSN full-length library NORMALIZATION oil accumulation Sesamue indicum Zhongzhi 14 cdna library switching mechanism SMARTTM
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Generation and Analysis of Expressed Sequence Tags(ESTs) from Muscle Full-Length cDNA Library of Wujin Pig 被引量:2
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作者 ZHAO Su-mei LIU Yong-gang +4 位作者 PAN Hong-bing ZHANG Xi GE Chang-rong JIA Jun-jing GAO Shi-zheng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2014年第2期378-386,共9页
Porcine skeletal muscle genes play a major role in determining muscle growth and meat quality. Construction of a full-length cDNA library is an effective way to understand the expression of functional genes in muscle ... Porcine skeletal muscle genes play a major role in determining muscle growth and meat quality. Construction of a full-length cDNA library is an effective way to understand the expression of functional genes in muscle tissues. In addition, novel genes for further research could be identified in the library. In this study, we constructed a full-length cDNA library from porcine muscle tissue. The estimated average size of the cDNA inserts was 1 076 bp, and the cDNA fullness ratio was 86.2%. A total of 1 058 unique sequences with 342 contigs (32.3%) and 716 singleton (67.7%) expressed sequence tags (EST) were obtained by clustering and assembling. Meanwhile, 826 (78.1%) ESTs were categorized as known genes, and 232 (21.9%) ESTs were categorized as unknown genes. 65 novel porcine genes that exhibit no identity in the TIGR gene index of Sus scrofa and 124 full-length sequences with unknown functions were deposited in the dbEST division of GenBank (accession numbers: EU650784-EU650788, GE843306, GH228978-GH229100). The abundantly expressed genes in porcine muscle tissue were related to muscle fiber development, energy metabolism and protein synthesis. Gene ontology analysis showed that sequences expressed in porcine muscle tissue contained a high percentage of binding activity, catalytic activity, structural molecule activity and motor activity, which involved mainly in metabolic, cellular and developmental process, distributed mainly in intracellular region. The sequence data generated in this study would provide valuable information for identifying porcine genes expressed in muscle tissue and help to advance the study on the structure and function of genes in pigs. 展开更多
关键词 muscle tissue full-length cdna library expressed sequence tag PIG
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Isolation and the Full-length Genome Sequencing of Subgroup J Avian Leukosis Virus ZH-08 Isolate Associated with Hemangioma 被引量:1
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作者 ZHANG Xiao-tao SHI Wei-wei LIU Hong-bo ZHANG He-nan LIAO Ming XIN Chao-an CAO Wei-sheng 《畜牧兽医学报》 CAS CSCD 北大核心 2011年第B12期51-56,共6页
A subgroup J avian leukosis virus (AVL-J), designated as ZH-08, was isolated from a breeder flock in Guangdong province with a novel hemangioma case. The identification results of ELISA test, PCR and immunofluoresecen... A subgroup J avian leukosis virus (AVL-J), designated as ZH-08, was isolated from a breeder flock in Guangdong province with a novel hemangioma case. The identification results of ELISA test, PCR and immunofluoresecence assay (IFA) specific for ALV-J were all positive. Based on the public full-length proviral genome sequence of ALV-J prototype strain HPRS-103, three pairs of primers were synthesized. The full-length proviral genome sequence of ZH-08 isolate is 7597 bp, which has a little difference with that of published full-length genome sequences, but its organization corresponds with typical retroviral genome structure; and known oncogenes were not included in its genome. According to the gp85 sequence comparison of ZH-08 isolate with those of the other reference strains in China and abroad, the highest similarity (93.7%) was with the YZ9901 isolate. Phylogenetic analysis, based on the gp85 gene, showed that the ZH-08 isolated here had the closest linkage to the SD07LK1 isolate. This study provides the basis for the biological characterization and pathogenesis research of the ZH-08 isolate. 展开更多
关键词 J亚群禽白血病病毒 全基因组测序 血管 全基因组序列 酶联免疫吸附试验 ALV-J 聚合酶链反应 GP85基因
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A NEW METHOD TO CONSTRUCT A FULL-LENGTH cDNA LIBRARY OF HUMAN NORMAL BLADDER TISSUE
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作者 成瑜 李旭 +2 位作者 陈葳 杨玉琮 赵乐 《Journal of Pharmaceutical Analysis》 SCIE CAS 2003年第2期173-175,188,共4页
Objective Using template switch mechanism at the 5’ end of mRNA technique (SMART) to construct a full length cDNA library of human normal bladder tissue. Methods The novel procedures used the template switchin... Objective Using template switch mechanism at the 5’ end of mRNA technique (SMART) to construct a full length cDNA library of human normal bladder tissue. Methods The novel procedures used the template switching activity of powerscript reverse transcriptase to synthesize and anchor first strand cDNA in one step. Following reverse transcription, 5 cycles of PCR were performed using a modified oligo(dT) primer and an anchor primer to enrich the full length cDNA population with 1.0 g human normal bladder poly(A) + RNA, then double strand cDNA was synthesized. After digestion with sfiI and size fractionation by CHROMA SPIN 400 columns, double strand cDNA was ligated into λ TripIEx 2 vector and was packaged. We determined the titer of the primary library and the percentage of recombinant clones and finally amplified the library. Results The titer of the cDNA library constructed was 2.1×10 6 pfu·mL -1 , and the amplified cDNA library was 6×10 11 pfu·mL -1 , the percentage of recombination clones was 99%. Conclusion Using SMART technique helps us to construct full length cDNA library with high efficiency and high capacity which lays solid foundation for screening target genes of bladder diseases with probes and antibodies. 展开更多
关键词 human normal bladder tissue cdna library full length λTripIEx 2
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An Introduction to China Rice Functional Genomics Program
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作者 Yong-biao XUE, Jing-liu ZHANG, Da LUO, Jia-yang LI, Yao-guang LIU, Hong-wei XUE, Kang CHONG, Hai HUANG, Guo-hua LIANG, Zhi-hong XU(1. Institute of Genetics and Developmental Biology, The Chinese Academy of Sciences,Beijing 100080, China 2. Institute of Plant Physiology and Ecology, TheChinese Academy of Sciences, Shanghai 200032, China +4 位作者 3. South China AgriculturalUniversity, Guangzhou 510642, China 4. Institute of Botany, The Chinese Academy ofScience, Beijing 100093, China 5. Yanzhou University, Yangzhou 225009, China 6. The Chinese Academy of Sciences,Beijing 100864 Beijing University,Beijing 100871, China) 《棉花学报》 CSCD 北大核心 2002年第S1期40-40,共1页
To discover genes essential for agronomicperformances of crops we initiated a program onrice functional genomics of important agronomictraits in rice in 1999.The program was fundedby the Ministry of Science and Techno... To discover genes essential for agronomicperformances of crops we initiated a program onrice functional genomics of important agronomictraits in rice in 1999.The program was fundedby the Ministry of Science and Technology 展开更多
关键词 genomicS crops genomicS AGRONOMIC initiated FLOWERING FERTILITY MUTANT GENETICS length
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Cloning and Sequence Analysis of the Full-Length Genome of Japanese encephalitis virus Strain SXBJ07 Isolated from Swine
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作者 WANG Wei-hua,ZHANG Yan-ming,XU Xin-gang,XING Fu-shan and TANG Qing-hai College of Veterinary Medicine,Northwest A&F University,Yangling 712000,P.R.China 《Agricultural Sciences in China》 CSCD 2009年第11期1392-1402,共11页
A virus strain, showing cytopathic effect in BHK-21 cell, was isolated from swine brains in Shaanxi Province, China, in 2007. The isolate was confirmed as Japanese encephalitis virus (JEV) by immunofluorescence ass... A virus strain, showing cytopathic effect in BHK-21 cell, was isolated from swine brains in Shaanxi Province, China, in 2007. The isolate was confirmed as Japanese encephalitis virus (JEV) by immunofluorescence assay (IFA) and reverse transcription polymerase chain reaction (RT-PCR), and named SXBJ07. The complete nueleotide and deduced amino acid sequences of the JEV strain SXBJ07 were determined. Its single open reading frame has a total of 3 432 amino acid residues. An extensive E gene based phylogenetic analysis was performed, the result showed that SXBJ07 strain belongs to genotype I. Comparison of the SXBJ07 genomic sequence with those of the 24 fully sequenced JEV strains in published databases showed nucleotide homology ranging from 99.0 to 83.7%; amino acid homology ranged from 99.8 to 94.8%. Compared SXBJ07 with SA14-14-2 strain, the current live vaccine strain in China, the homology of amino acid in envelope gene was 97.0%; and there were amino acid substitutions in 13 sites of the active domains of E protein (E1-E411). 展开更多
关键词 Japanese encephalitis virus (JEV) full-length genome sequence analysis
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端粒长度与10种常见肌肉骨骼疾病的关系孟德尔随机化分析
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作者 罗伟东 蒲彬 +3 位作者 古鹏 黄枫 郑晓辉 陈福洪 《中国组织工程研究》 CAS 北大核心 2025年第3期654-660,共7页
背景:多项观察性研究表明,端粒长度与肌肉骨骼疾病之间存在潜在的关联,然而它们之间的潜在机制仍不清楚。目的:利用两样本孟德尔随机化分析来探索端粒长度与肌肉骨骼疾病之间的遗传因果关系。方法:从英国生物银行中获得端粒长度的全基... 背景:多项观察性研究表明,端粒长度与肌肉骨骼疾病之间存在潜在的关联,然而它们之间的潜在机制仍不清楚。目的:利用两样本孟德尔随机化分析来探索端粒长度与肌肉骨骼疾病之间的遗传因果关系。方法:从英国生物银行中获得端粒长度的全基因组关联研究汇总数据。从FinnGen财团中获得了关于10种常见肌肉骨骼疾病(骨坏死、骨髓炎、骨质疏松、类风湿关节炎、腰痛、椎管狭窄、痛风、肩周炎、强直性脊柱炎和下肢深静脉血栓)的全基因组关联研究汇总数据。使用逆方差加权、孟德尔随机化-Egger和加权中位数方法评估端粒长度与10种肌肉骨骼疾病的因果关系,逆方差加权作为主要的孟德尔随机化分析方法,并进行敏感性分析探讨结果稳健性。结果与结论:①逆方差加权法结果表明,遗传预测的端粒长度与类风湿关节炎(OR=0.78,95%CI:0.64-0.95,P=0.015)和骨坏死(OR=0.56,95%CI:0.36-0.90,P=0.016)风险之间存在负向因果关系,但未发现端粒长度与其他8种肌肉骨骼疾病之间存在因果关系(P均>0.05)。②敏感性分析结果表明因果关系稳健,孟德尔随机化-Egger截距分析未检测到潜在的水平多效性(P均>0.05)。③此项孟德尔随机化研究支持端粒长度对类风湿关节炎和骨坏死的保护作用的结论,然而,未来将需要更多的基础和临床研究来验证。 展开更多
关键词 端粒长度 肌肉骨骼疾病 孟德尔随机化 全基因组关联研究 单核苷酸多态性 因果关系 工具变量
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猪瘟病毒兔化弱毒(HCLV)疫苗株基因组全长cDNA的克隆与序列分析 被引量:9
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作者 余兴龙 涂长春 +3 位作者 徐兴然 张茂林 李作生 于师宇 《高技术通讯》 EI CAS CSCD 2003年第5期38-42,共5页
根据猪瘟病毒基因组的序列设计合成了覆盖HCLV基因组全部序列的5对引物,通过RT-PCR从感染家兔脾脏中扩增获得了包含HCLV(bog cholera lapinized virus,HCLV)基因组全序列的5个cDNA片段,并将它们分别克隆至pGEM-T vector中。然后将这5个c... 根据猪瘟病毒基因组的序列设计合成了覆盖HCLV基因组全部序列的5对引物,通过RT-PCR从感染家兔脾脏中扩增获得了包含HCLV(bog cholera lapinized virus,HCLV)基因组全序列的5个cDNA片段,并将它们分别克隆至pGEM-T vector中。然后将这5个cDNA片段按它们在HCLV基因组上的位置从5’端至3’端的顺序依次通过酶切连接,一并克隆到pPoly2载体中得到HCLV基因组全长cDNA的质粒pPOHCLV。序列分析表明,HCLV基因组长度共12310bp,有一个大的ORF,编码一3898个氨基酸的多聚蛋白。其5'-NCR和3’-NCR长度分别是374nt和239nt。与非兔化毒株相比较,HCLV基因组在12133nt处有12个碱基的插入CTTTTTTCTTTT,该序列可能是病毒在适应兔体增殖过程中形成的。基于基因组全序列及其所推导的氨基酸序列的同源性分析表明,毒株的亲缘关系的远近与它们的毒力之间并没有相关性。 展开更多
关键词 猪瘟病毒 猪瘟兔化弱毒 基因组 疫苗株 HCLV cdna 全长序列 克隆 同源性
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庚型肝炎病毒基因组全长cDNA的拼接及克隆 被引量:13
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作者 朱分禄 戚中田 +2 位作者 任浩 宋燕斌 邵力 《第二军医大学学报》 CAS CSCD 北大核心 1998年第4期301-306,共6页
目的:构建GB病毒C/庚型肝炎病毒(GBV-C/HGV)基因组全长cDNA克隆。方法:以覆盖GBV-C/HGV分离株HGV-Iw全长基因组且互相重叠的5个基因片段Iw5,Iwq2,Iwh6,Iw3和Iw3为起始材料... 目的:构建GB病毒C/庚型肝炎病毒(GBV-C/HGV)基因组全长cDNA克隆。方法:以覆盖GBV-C/HGV分离株HGV-Iw全长基因组且互相重叠的5个基因片段Iw5,Iwq2,Iwh6,Iw3和Iw3为起始材料,采用重叠延伸PCR拼接和连接酶连接的方法,构建GBV-C/HGV基因组全长cDNA克隆。结果:GBV-C/HGV基因组全长cDNA克隆的酶切图谱与预期一致;核苷酸序列分析显示,克隆的GBV-C/HGV基因组全长cDNA的核苷酸及推测的氨基酸序列与原HGV-Iw序列一致。结论:GBV-C/HGV基因组全长cDNA的克隆已经构建成功。该克隆的构建,为深入研究GBV-C/HGV的致病性及致病机制、复制及转录和翻译机制。 展开更多
关键词 全长cdna 拼接 克隆 庚型肝炎病毒 基因组
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猪乙型脑炎病毒SH0601株基因组序列分析及全长cDNA的构建 被引量:8
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作者 郑浩 余丹丹 +2 位作者 孙志 高飞 袁世山 《中国人兽共患病学报》 CAS CSCD 北大核心 2008年第2期140-145,共6页
目的对猪源乙型脑炎病毒SH0601株进行全序列测定与分析,以了解该株病毒的遗传特征,并构建全长cDNA克隆。方法将猪源JEV SH0601株基因组RNA分5段进行RT-PCR并克隆入pCRⅡ-Blunt-TOPO和pBluescript载体中,测定了全长cDNA序列。根据cDNA克... 目的对猪源乙型脑炎病毒SH0601株进行全序列测定与分析,以了解该株病毒的遗传特征,并构建全长cDNA克隆。方法将猪源JEV SH0601株基因组RNA分5段进行RT-PCR并克隆入pCRⅡ-Blunt-TOPO和pBluescript载体中,测定了全长cDNA序列。根据cDNA克隆酶切图谱分析,将5个片段先后克隆入pBluescript中构建JEV全长基因组cDNA。根据E蛋白基因序列对SH0601株进行了遗传分型。结果序列分析表明:SH0601株基因组全长10977nt,与GenBank中的Beijing-1、P3、SA14和SA14-14-2株基因组全长相比,在3′NTR的10701nt处多一碱基"G",与上述4株病毒的全长核苷酸的同源率都达97.2%以上,而与国外猪源分离株KV1899和JEV/sw/Mie/41/2002全长核苷酸的同源率仅达88.7%,但SH0601株与6株病毒的全长氨基酸同源率均达97.0%以上。以E蛋白基因序列对SH0601作进化分析,该分离株属基因型Ⅲ型。结论SH0601株属于乙型脑炎病毒基因Ⅲ型。在高拷贝质粒pBluescript中可构建稳定的乙型脑炎病毒全长cDNA克隆。 展开更多
关键词 乙型脑炎病毒 病毒基因组 全长cdna克隆
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与绵羊KAP6-1相似的6个绒山羊全长cDNA的克隆与序列分析 被引量:37
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作者 尹俊 扈庭茂 +3 位作者 李金泉 张春兰 郭志成 周欢敏 《Acta Genetica Sinica》 SCIE CAS CSCD 北大核心 2004年第5期502-507,共6页
用SMARTTM技术构建了绒山羊体侧部皮肤组织的cDNA文库 ,随机挑选克隆提取质粒 ,用M13正向测序引物对插入片段进行测序 ,得到 6 36个cDNA序列。与GenBank数据库比对 ,有 4 1个序列与绵羊角蛋白关联蛋白(KeratinAssociatedProtein ,KAP6 ... 用SMARTTM技术构建了绒山羊体侧部皮肤组织的cDNA文库 ,随机挑选克隆提取质粒 ,用M13正向测序引物对插入片段进行测序 ,得到 6 36个cDNA序列。与GenBank数据库比对 ,有 4 1个序列与绵羊角蛋白关联蛋白(KeratinAssociatedProtein ,KAP6 1)cDNA高度同源。 4 1个序列可归为 6个不同的cDNA ,GenBank登陆号分别为AY310 74 9、AY310 75 0、AY310 75 1、AY310 75 2、AY310 75 3和AY310 75 4。与绵羊KAP6 1基因组基因比较 ,推测这 6个序列为全长cDNA ,分别为编码 82、84、71、71、83和 83个氨基酸的碱性蛋白质 ,甘氨酸和酪氨酸的含量大于 6 0 % ,它们之间核苷酸序列的一致性大于 5 5 4 % ,读码框氨基酸序列的一致性大于 79 8%。与其他物种KAP6s比较 ,绒山羊的 6个cDNA和绵羊的KAP6 1cDNA的序列一致性最高 ,为 81 9%~ 98 8% ,不同物种KAP6 1之间氨基酸序列一致性大于 5 0 %。 展开更多
关键词 绒山羊 KAP6-1 全长cdna
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cDNA文库构建策略及其分析研究进展 被引量:81
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作者 晏慧君 黄兴奇 程在全 《云南农业大学学报》 CAS CSCD 2006年第1期1-6,共6页
cDNA文库构建和筛选是基因克隆的重要方法之一,它是目前发现新基因和研究基因功能的基本工具。从cDNA文库中可以筛选到目的基因,并直接用于该基因的表达。经典cDNA文库存在克隆的片段短等缺点,而全长cDNA文库则能提供完整的mRNA信息,从... cDNA文库构建和筛选是基因克隆的重要方法之一,它是目前发现新基因和研究基因功能的基本工具。从cDNA文库中可以筛选到目的基因,并直接用于该基因的表达。经典cDNA文库存在克隆的片段短等缺点,而全长cDNA文库则能提供完整的mRNA信息,从而克隆cDNA全长;对文库进行均一化处理即均一化cDNA文库,可以增加克隆低丰度mRNA的机会;差减cDNA文库在研究生物体某一时期基因差异表达上具有重要的意义;改进的固相cDNA很大程度上提高了建库的效率和质量。本文以阐述基本原理为主,介绍几种近年来常用的cDNA构建的方法及其优缺点。 展开更多
关键词 cdna文库 cdna全长 基因克隆 均一化cdna文库 差减cdna文库 固相cdna文库 快速扩增cdna末端(RACE)
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内蒙古绒山羊毛囊兴盛期皮肤cDNA文库的构建及KAP6-2 cDNA的克隆 被引量:9
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作者 尹俊 扈庭茂 +3 位作者 李金泉 张春兰 郭志成 周欢敏 《Zoological Research》 CAS CSCD 北大核心 2004年第2期166-171,共6页
用SMART技术构建了绒山羊 (Caprahircus)毛囊兴盛期皮肤组织的cDNA质粒文库。TrizolReagent(GIBCO/BRL)分离RNA后 ,用Oligotex (QIAGEN)提取mRNA ;以锚定引物反转录合成cDNA第一链作为模板 ,以 2个锚定引物 ,用长链PCR扩增全长cDNA双链... 用SMART技术构建了绒山羊 (Caprahircus)毛囊兴盛期皮肤组织的cDNA质粒文库。TrizolReagent(GIBCO/BRL)分离RNA后 ,用Oligotex (QIAGEN)提取mRNA ;以锚定引物反转录合成cDNA第一链作为模板 ,以 2个锚定引物 ,用长链PCR扩增全长cDNA双链。CHROMASPIN 4 0 0去除小片段后用SfiⅠ酶切 ,连接到SfiⅠ消化的pBluescriptⅡSK (带有SfiⅠA和B两个位点 )质粒载体中 ,转化E .coli 5α ,库容量为 1 8×10 5clones。随机挑选克隆提取质粒 ,5′测定插入片段的核苷酸序列 ,与GenBank数据库比对后发现 1个KAP6 2cDNA ,序列号为AY316 15 8。绒山羊KAP6 2由 6 75个核苷酸组成 ,编码 96个氨基酸 ,具有高甘氨酸 展开更多
关键词 内蒙古绒山羊 毛囊兴盛期 皮肤组织 cdna文库 核苷酸序列
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油茶SAD基因的全长cDNA克隆及生物信息学分析 被引量:42
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作者 张党权 谭晓风 +4 位作者 陈鸿鹏 曾艳玲 蒋瑶 李魏 胡芳名 《林业科学》 EI CAS CSCD 北大核心 2008年第2期155-159,共5页
As a high-grade edible oil tree native in China,tea-oil tree(Camellia oleifera)has the oil-yielded rate of about 55% from its kernel.The recent researches suggested that tea-oil would be one of the best vegetable oils... As a high-grade edible oil tree native in China,tea-oil tree(Camellia oleifera)has the oil-yielded rate of about 55% from its kernel.The recent researches suggested that tea-oil would be one of the best vegetable oils,and even be better than olive oil with its abundant unsaturated fatty acids including 82.6% oleic acid.Stearoyl-ACP desaturase(SAD)is a key enzyme that catalyzes saturated fatty acids(C18∶0)bonded to ACP(Acyl carrier protein)and dehydrogenates the fatty acids into oleic acids,and hence controls the content of oleic acid and the proportion between saturated fatty acids and unsaturated fatty acids.With our previous acquisition of three cDNAs and ESTs of C.oleifera SAD(CoSAD)gene,5’RACE technology was used to obtain the full-length cDNA of CoSAD gene from the nearly matured C.oleifera seed.The comprehensive bioinformatic analyses including sequence characteristics of DNA and amino acid,multi-sequence aligning,identity and homology,molecular clustering,protein physicochemical properties,and protein structural prediction and characteristics were performed.The results may provide the theoretical and material elements for application of CoSAD gene and genetic improvement on other oil plants. 展开更多
关键词 油茶 硬脂酰-ACP脱饱和酶(SAD) 油酸 全长cdna 生物信息学
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cDNA末端快速扩增技术及其应用 被引量:18
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作者 王少丽 盛承发 乔传令 《遗传》 CAS CSCD 北大核心 2004年第3期419-423,共5页
cDNA末端快速扩增(RACE)技术是一种快速获得cDNA的3’和5’端的方法。从RACE的原理出发,指出其技术本身存在的优缺点,阐述了RACE操作中须注意和不容忽视的技术要点,并对前人对RACE技术所做的改进加以总结,最后对RACE技术的应用前景给予... cDNA末端快速扩增(RACE)技术是一种快速获得cDNA的3’和5’端的方法。从RACE的原理出发,指出其技术本身存在的优缺点,阐述了RACE操作中须注意和不容忽视的技术要点,并对前人对RACE技术所做的改进加以总结,最后对RACE技术的应用前景给予了展望。 展开更多
关键词 RACE 全长cdna 应用
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全长cDNA文库的构建方法 被引量:22
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作者 董志敏 张宝石 +2 位作者 关荣霞 常汝镇 邱丽娟 《中国农学通报》 CSCD 2006年第2期51-55,共5页
全长cDNA文库的构建是进行功能基因组研究的一种经济、快速、有效的途径,克服了传统cDNA文库的缺点,节省了基因克隆和功能鉴定的时间,促进了功能基因组的研究进程。目前有6种构建全长cDNA文库的方法,包括Oligo-Capping、CAPture、SMART... 全长cDNA文库的构建是进行功能基因组研究的一种经济、快速、有效的途径,克服了传统cDNA文库的缺点,节省了基因克隆和功能鉴定的时间,促进了功能基因组的研究进程。目前有6种构建全长cDNA文库的方法,包括Oligo-Capping、CAPture、SMART、CAP-trapper、CapSelect、CAP-jumping。这几种方法在起始材料用量、文库构建技术、实验操作复杂程度和文库构建质量等方面构存在不同程度的优缺点,但综合比较而言,SMART和CAP-trapper这两种方法比较有实际应用价值。SMART法适于简单、快速地构建一般质量的cDNA文库,如果采用Little-cycleSMART和Large-sizeSMART这两种改进技术,所建文库质量也非常好;CAP-trapper法虽然对实验技术要求较高,实验过程复杂,但所建文库全长率高,冗余性低,建库效率高,是目前构建高质量文库最好的方法。 展开更多
关键词 全长cdna 5’帽子结构 cdna文库的构建
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