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Expressed sequence tags (ESTs) analysis of the ripening Vitis amurensis cv. Shuang Hong berry skins 被引量:1
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作者 JI Xiang-Nan LI Feng +3 位作者 YANG Cheng-Jun LI Bo WANG Jun ZHANG Wen 《Journal of Forestry Research》 SCIE CAS CSCD 2013年第3期495-502,共8页
Vitis amurensis is a valuable resource for wine production. Ripening of the grape berry is the key phase which determines the com- position of wine. To better understand the gene expression that manifest in V. amurens... Vitis amurensis is a valuable resource for wine production. Ripening of the grape berry is the key phase which determines the com- position of wine. To better understand the gene expression that manifest in V. amurensis berry skins during the ripening, cDNA library of V. amurensis berry skins was constructed. A total of 935 high quality ex- pressed sequence tags (ESTs) were obtained from the library. These ESTs represent 636 unigenes, including 108 contigs and 528 singletons. The EST analysis was performed and genes were assigned to functional categories according to their primary BLAST match. Of these 25.35% were involved with metabolism, 6.27% with cell rescue and defense, 6.84% energy, 11.68% protein synthesis, 18.8% protein activity regula- tion, 11.11% cell structure, 7.98% transport, 6.27% transcription and the remaining 5.7% were signal transduction. The generated ESTs were characterized by the gene ontology analysis and were categorized ac- cording to its cellular component, molecular function and biological process. In the cDNA library, some genes are relevant to the biosynthesis of anthocyanins, while some genes are related to grape berry maturation. 展开更多
关键词 Vitis amurensis expressed sequence tags ests) cDNAlibrary unigene annotation gene ontology
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基于EST-SSR标记的沙棘品种鉴定及指纹图谱构建
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作者 赵雨欣 张哲文 +5 位作者 考惠霞 孙永江 辛智鸣 赵喆 董树斌 程瑾 《生态与农村环境学报》 CAS CSCD 北大核心 2024年第3期374-385,共12页
以沙棘(Hippophae rhamnoides)优良品种“实优1号”为材料,对其叶片进行转录组测序,利用微卫星识别软件(microsatellite identification tool,MISA)和Primer 3(version 2.3.4)对获得的序列进行简单重复序列(simple sequence repeat,SSR... 以沙棘(Hippophae rhamnoides)优良品种“实优1号”为材料,对其叶片进行转录组测序,利用微卫星识别软件(microsatellite identification tool,MISA)和Primer 3(version 2.3.4)对获得的序列进行简单重复序列(simple sequence repeat,SSR)位点挖掘和引物设计,以收集的42份沙棘品种为研究材料,开展聚合酶链式反应(PCR)和毛细管电泳检测,旨在开发一套多态性高、稳定性好和通用性强的表达序列标签微卫星(express sequence tags from simple sequence repeat,EST-SSR)引物,构建沙棘指纹图谱,从而实现沙棘品种的快速准确鉴定,并对沙棘品种间亲缘关系进行分析。“实优1号”转录组测序共获得6196个SSR位点,其中,重复基元类型为182种,SSR基序长度主要分布在10~21 bp区间,占全部SSR的81.58%,主要SSR重复类型为单核苷酸重复(48.72%)、二核苷酸重复(22.68%)和三核苷酸重复(18.85%)。利用筛选出的28对引物在42份沙棘品种中共检测出193个等位基因,等位基因数(Na)、有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)和Shannon信息指数(I)等遗传多样性参数的均值分别为6.964、3.495、0.617、0.671、0.623和1.384。UPGMA聚类分析表明,42份沙棘品种间的遗传相似性系数为0.601~0.990,当遗传相似性系数为0.694时,供试品种可分为2组;当遗传相似性系数约为0.7402时,供试品种可分为3组。优选6对引物构建指纹图谱,可以实现沙棘品种的快速准确鉴定。该研究可为沙棘的良种鉴定、指纹图谱构建以及遗传多样性和亲缘关系分析等提供分子水平的理论基础和数据支撑。 展开更多
关键词 沙棘 表达序列标签微卫星 指纹图谱 遗传多样性
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Primarily screening and analyzing ESTs differentially expressed in rats' primary liver cancer 被引量:9
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作者 Dandan Liu Lijuan Zhi +9 位作者 Mingxia Ma Dan Qiao Meijuan Wang Yawei Wang Baijie Jin Anqi Li Guting Liu Yiqing Zhang Yanyan Song Hongxu Zhang 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2013年第1期71-78,共8页
Objective: To screen and analyze key express sequence tags (ESTs) which were differentially displayed in every period of SD rats' primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Met... Objective: To screen and analyze key express sequence tags (ESTs) which were differentially displayed in every period of SD rats' primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Methods: Using diethylnitrosamine (DENA) as a cancerigenic agent, animal models with different phases of primary hepatic cancer were constructed in SD rats. Rats were respectively sacrificed at d 14, d 28, d 56, d 77, d 105 and d 112 after the rats received DENA by gavage, then the livers were harvested. One part of the livers was classified according to their pathological changes, while the other was reserved for molecular mechanism studies on hepatocarcinogenesis. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique (DDRT-PCR). After the fragments were sequenced, bioinformatics were .used to analyze the results. Results: Twelve differentially expressed cDNA fragments were obtained. Nine fragments had the homology with known cDNA clones, especially EST-7 was similar to BN/SsNHsdMCW mitochondrion gene and the identity was 100% which suggested EST-7 may be the part of BN/SsNHsdMCW mitochondrion gene. In contrast, other three fragments (EST-1, EST-3 and EST-5) had extremely low identity to any genes registered in GENBANK databases. Conclusions: BN/SsNHsdMCW mitochondrion gene was expressed in different periods of hepatocarcinogenesis. Moreover, EST-I, EST-3 and EST-5 were suggested to contribute to the development of rat hepatocarcinogenesis, and thus may be candidates of new targets of oncogenes or cancer suppressor genes. 展开更多
关键词 Animal models of primary liver cancer DDRT-PCR (differential display reverse transcription PCR) ests express sequence tags) mitochondrion gene
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Generation and analysis of expressed sequence tags from the salt-tolerant eelgrass species, Zostera marina 被引量:5
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作者 KONG Fanna ZHOU Yang +2 位作者 SUN Peipei LIU Limin MAO Yunxiang 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2013年第8期68-78,共11页
Zostera marina, a monocotyledonous angiosperm, is one of the most important seagrass species. To inves- tigate the salt-tolerance mechanism and discover salt-tolerant genes in Z. marina, a cDNA library was con- struct... Zostera marina, a monocotyledonous angiosperm, is one of the most important seagrass species. To inves- tigate the salt-tolerance mechanism and discover salt-tolerant genes in Z. marina, a cDNA library was con- structed. Single-pass sequencing of the 5' ends of 4 081 clones yielded 4 002 high quality expressed sequence tags (ESTs), which were assembled into 241 contigs and 1 673 singletons, representing 1 914 unigenes. The average length of the ESTs was 582 bp, with sizes ranging from 100-1 500 bp. Basic Local Alignment Search Tool (BLASTX) analysis revealed that 1 664 unigenes had significant homology to known genes in the Na- tional Center for Biotechnology Information (NCBI) non-redundant (nr) database (E-value≤5-10). Among them, the two most abundant genes encoded metallothionein (157 ESTs) and chlorophyll a/b-binding pro- tein (38 ESTs), accounting for 7.1% and 1.7% of the total ESTs, respectively. Using Kyoto Encyclopedia of Genes and Genomes (KEGG), 1 462 unigenes were assigned to 1 161 pathways (E-value≤5-10). A total of 938 unigenes were assigned Gene Ontology (GO) terms based on the GO hierarchy analysis, and InterProScan searches recognized 1 003 InterPro families. Three genes for metallothionein in Z. marina that belonged to Class II was identified. Results of this study will improve understanding of the molecular mechanisms of saline tolerance in Z. marina. 展开更多
关键词 expressed sequence tags ests) gene function METALLOTHIONEIN salt-tolerant genes Zostera marina
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Analysis of Expressed Sequence Tags from Liver Tissue in Swine 被引量:1
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作者 LI Ning, ZHAO Zhi-hui, LIU Zhao-Hang, ZHAO Xing-bo, LIAN Zhen-xing and WU Chang-xin(National Laboratory for Agro-biotechnology, China Agricultural University, Beijing 100094 , P.R.China College of Animal Science and Technology, China Agricultural University , Beijing 100094 , P. R . China ) 《Agricultural Sciences in China》 CAS CSCD 2002年第9期1050-1053,共4页
In order to study the expression of function gene and its effect on metabolic control and other physiological function in liver, 438 expressed sequence tags (ESTs) were determined, which were from a cD-NA library of p... In order to study the expression of function gene and its effect on metabolic control and other physiological function in liver, 438 expressed sequence tags (ESTs) were determined, which were from a cD-NA library of porcine liver tissue. The results showed that the nucleotide sequences of 186 ESTs have already presented in GenBank database, and 37 ESTs could be found the homology with human and other species, while the others were not identified. 45 full length insertion of the clones randomly isolated from cDNA library were also completely sequenced with different size, and the results showed that 19 of them were function-known genes, 11 had no open reading frame ( ORF )at all and 15 had ORF but the function were not elucidated yet. 展开更多
关键词 SWINE LIVER cDNA library expressed sequence tags(ests)
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Analysis of expressed sequence tags from the Ulva prolifera (Chlorophyta) 被引量:4
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作者 牛建峰 胡海岩 +3 位作者 胡松年 王广策 彭光 孙松 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第1期26-36,共11页
In 2008,a green tide broke out before the sailing competition of the 29th Olympic Games in Qingdao.The causative species was determined to be Enteromorpha prolifera (Ulva prolifera O.F.Müller),a familiar green ma... In 2008,a green tide broke out before the sailing competition of the 29th Olympic Games in Qingdao.The causative species was determined to be Enteromorpha prolifera (Ulva prolifera O.F.Müller),a familiar green macroalga along the coastline of China.Rapid accumulation of a large biomass of floating U.prolifera prompted research on different aspects of this species.In this study,we constructed a nonnormalized cDNA library from the thalli of U.prolifera and acquired 10 072 high-quality expressed sequence tags (ESTs).These ESTs were assembled into 3 519 nonredundant gene groups,including 1 446 clusters and 2 073 singletons.After annotation with the nr database,a large number of genes were found to be related with chloroplast and ribosomal protein,GO functional classification showed 1 418 ESTs participated in photosynthesis and 1 359 ESTs were responsible for the generation of precursor metabolites and energy.In addition,rather comprehensive carbon fixation pathways were found in U.prolifera using KEGG.Some stress-related and signal transduction-related genes were also found in this study.All the evidences displayed that U.prolifera had substance and energy foundation for the intense photosynthesis and the rapid proliferation.Phylogenetic analysis of cytochrome c oxidase subunit I revealed that this green-tide causative species is most closely affiliated to Pseudendoclonium akinetum (Ulvophyceae). 展开更多
关键词 表达序列标签 绿藻门 细胞色素C氧化酶 ests cDNA文库 系统发育分析 光合作用 est序列
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Characterization of genic microsatellite markers derived from expressed sequence tags in Pacific abalone (Haliotis discus hannai) 被引量:1
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作者 李琪 束婧 +3 位作者 赵翠 刘士凯 孔令锋 郑小东 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第1期46-54,共9页
Simple sequence repeat (SSR) markers were developed from the expressed sequence tags (ESTs) of Pacific abalone (Haliotis discus hannai).Repeat motifs were found in 4.95% of the ESTs at a frequency of one repeat every ... Simple sequence repeat (SSR) markers were developed from the expressed sequence tags (ESTs) of Pacific abalone (Haliotis discus hannai).Repeat motifs were found in 4.95% of the ESTs at a frequency of one repeat every 10.04 kb of EST sequences,after redundancy elimination.Seventeen polymorphic EST-SSRs were developed.The number of alleles per locus varied from 2-17,with an average of 6.8 alleles per locus.The expected and observed heterozygosities ranged from 0.159 to 0.928 and from 0.132 to 0.922,respectively.Twelve of the 17 loci (70.6%) were successfully amplified in H.diversicolor.Seventeen loci segregated in three families,with three showing the presence of null alleles (17.6%).The adequate level of variability and low frequency of null alleles observed in H.discus hannai,together with the high rate of transportability across Haliotis species,make this set of EST-SSR markers an important tool for comparative mapping,marker-assisted selection,and evolutionary studies,not only in the Pacific abalone,but also in related species. 展开更多
关键词 表达序列标签 等位基因数 微卫星标记 皱纹盘鲍 太平洋 鲍鱼 简单重复序列 SSR标记
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Analysis of Simple Sequence Repeats Information from Floral Expressed Sequence Tags Resources of Papaya (<i>Carica papaya</i>L.)
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作者 Priyanka Priyanka Dileep Kumar +2 位作者 Anurag Yadav Kusum Yadav U. N. Dwivedi 《American Journal of Plant Sciences》 2017年第9期2315-2331,共17页
Papaya (Carica papaya L.) is one of the most economically, medicinally and nutritionally important tropical fruit crops. Expressed sequence tags (ESTs) derived simple sequence repeat (SSR) markers are more valuable as... Papaya (Carica papaya L.) is one of the most economically, medicinally and nutritionally important tropical fruit crops. Expressed sequence tags (ESTs) derived simple sequence repeat (SSR) markers are more valuable as they are derived from conserved genic portion. Development of EST-SSRs markers through in silico approach is cheaper, less time consuming and labour-intensive. In this study, we aimed to mine SSRs and developed EST-SSR primers from papaya floral ESTs. A total of 75,846 papaya floral ESTs were downloaded from public database National Centre for Biotechnology Information (NCBI). A total of 26,039 floral unigenes (7961 contigs and 18,078 singletons) were generated after assembly of these ESTs. From these floral unigenes, 433,782 perfect SSRs, 204,968 compound SSRs and 6061 imperfect SSRs were mined, respectively. In perfect SSRs, mononucleotide repeats were most abundant (94.7%) followed by tri- (3.1%) and di-nucleotide repeats (1.7%). The frequencies of tetra-, hexa- and penta-nucleotide repeats accounted for only (0.17%), (0.04%) and (0.03%), respectively. In mononucleotide repeats, the most abundant motif was A/T (69.3%) and in di- and tri-nucleotide repeats were AG/CT (61%) and AAG/CTT (31%), respectively. In imperfect SSRs, mononucleotide repeats (56.5%) were most abundant. 176 different types of motifs were identified. A total of 3807 primer pairs for floral papaya ESTs were successfully designed. These developed EST-SSR primers are being used for the genetic improvement of papaya such as study of cross-transferability across genera/species, evaluation of genetic diversity, and identification of sex-specific markers. These EST derived SSRs can also be used in filling gaps in existing linkage maps in papaya. 展开更多
关键词 PAPAYA (Carica PAPAYA L.) In Silico Simple SEQUENCE Repeats expressed SEQUENCE tags (ests) SSR Mining est-SSR SSR Motifs Primer Pairs
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Identification of Differentially Expressed Genes in the Salivary Gand of Rhipicephalus haemaphysaloides by the Suppression Subtractive Hybridization Approach 被引量:1
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作者 XIANG Fei-yu ZHOU Yong-zhi ZHOU Jin-lin 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2012年第9期1528-1544,共17页
For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and f... For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization (SSH). A total of 247 female expression sequence tags (ESTs) and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5" and 3" ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5" and 3" ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 (57%) female ESTs and 125 (74%) male ESTs had similarity to GenBank sequences, and 32 (23%) female ESTs and 29 (23%) male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks. 展开更多
关键词 Rhipicephalus haemaphysaloides salivary gland suppression subtractive hybridization (SSH) cDNA library expression sequence tag est
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Screening the stage-specific expression gene from different germ cells of rat
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作者 Jia Mengchun(贾孟春) +7 位作者 Lou Lixia(娄利霞) Pei Kaiyan(裴开颜) Zhao Longmei(赵龙梅) Shi Xinquan(石心泉) 《生殖医学杂志》 CAS 2002年第z1期29-37,共9页
Objective: To screen the stage-specific expression genes from rat spermatogonia, pachytene spermatocytes and round spermatids. Methods: Highly purified spermatogonia were isolated from 9-day-old rats, pachytene sperma... Objective: To screen the stage-specific expression genes from rat spermatogonia, pachytene spermatocytes and round spermatids. Methods: Highly purified spermatogonia were isolated from 9-day-old rats, pachytene spermatocytes and round spermatids from adult rats by sedimentation velocity at unit gravity, using 2%-4% BSA gradient in DMEM/F12 medium. A mRNA differential display method was used for screening the stage-specific expression gene. Results: Nineteen differentially expressed cDNA fragments were obtained. After excluding the false positive cDNA fragments by dot blot, 13 cDNAs were selected to clone and sequence. To obtain longer cDNAs, six ESTs were used to screen the rat testis λ-zap II cDNA library. Two longer cDNA fragments, designated as LY21 and LM66, were obtained. The analysis with DNAMAN software indicated that LY21 had a long open reading frame coding 372 amino acids while LM66 had no long open reading frame. LY21 were highly homologous with hnRNP H1. To observe the expression patterns of LY21 gene in the testicular cells, we performed in situ hybridization on testis sections from adult rats. The LY21 gene expression was found in the spermatogonia and primary spermatocytes.Conclusion: This study indicated that LY21 gene was associated with spermatogenesis. Further studies will be needed to explore the function of LY21. 展开更多
关键词 SPERMATOGENESIS Differential DISPLAY method express SEQUENCE tag (est) GENE
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Effect of 4% glycerol and low aeration on result of expression in <i>Escherichia coli</i>of Cin3 and three <i>Venturia inaequalis</i>EST’s recombinant proteins
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作者 Taha H. Al-Samarrai William T. Jones +2 位作者 Dawn Harvey Christopher A. Kirk M. Templtone 《American Journal of Molecular Biology》 2013年第1期1-9,共9页
The phytopathogenic fungus Venturia inaequalis causes scab of apple. Once this fungus penetrates the plant surface, it forms a specialized body called a stroma between the inner cuticle surface and the epidermal cell ... The phytopathogenic fungus Venturia inaequalis causes scab of apple. Once this fungus penetrates the plant surface, it forms a specialized body called a stroma between the inner cuticle surface and the epidermal cell wall. A novel Venturia inaequalis 5704 (Cin3) and three expressed sequence tags (ESTs);38, 6987, and 4010 are strongly up-regulated in the early stages of infection. The CIN3 and three ESTs using two vectors pMAL-c2 and pET 21 were expressed in Escherichia coli. Recombinant proteins expression, solubility and yields were analyzed. 38, 5704 (Cin3) and 6987 re- combinant proteins were expressed in soluble form and while 4010 was expressed in inclusion bodies. Re- solution on native-PAGE, the recombinant proteins;38, 5704 (Cin3), 6987 were shown to be present in dimmer, tetramer and polymer. A method was de- veloped, consisting of induction of expression at va- rious temperatures, and using enriched broth with 4% glycerol together with slow shaking, led to a decrease in concentration of nascent polypeptide and production of soluble recombinant proteins of;38, 5704 (Cin3), 6987 and 4010. Resolution on native- PAGE, the recombinant proteins were shown to be present as monomer. 展开更多
关键词 Venturia inaequalis expressed Sequence tag (ests) Phytopathogenic FUNGUS APPRESSORIUM A Stroma
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美洲鲥脑转录组多态性EST-SSR的规模化开发与利用 被引量:1
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作者 于爱清 施永海 +1 位作者 徐嘉波 刘永士 《西北农林科技大学学报(自然科学版)》 CSCD 北大核心 2023年第11期1-12,共12页
【目的】挖掘美洲鲥脑组织转录组数据中的EST-SSR,规模化开发多态性较好的EST-SSR,为美洲鲥种质资源评估提供分子标记。【方法】采集2龄美洲鲥雌雄个体(各3尾)的脑组织,提取其RNA,构建cDNA文库,进行高通量转录组测序。用MISA软件进行脑... 【目的】挖掘美洲鲥脑组织转录组数据中的EST-SSR,规模化开发多态性较好的EST-SSR,为美洲鲥种质资源评估提供分子标记。【方法】采集2龄美洲鲥雌雄个体(各3尾)的脑组织,提取其RNA,构建cDNA文库,进行高通量转录组测序。用MISA软件进行脑组织转录组EST-SSR挖掘和特征分析,利用SSRMMD软件规模化开发多态性EST-SSR。从获得的多态性位点中随机选取20对4核苷酸重复的EST-SSR设计引物,对24尾1龄美洲鲥进行遗传多样性评估,验证这些多态性EST-SSR的应用效果。【结果】从雌性美洲鲥脑组织转录组189 428个非冗余基因中鉴定到117 751个EST-SSR,从雄性美洲鲥脑组织转录组185 419个非冗余基因中鉴定到114 809个EST-SSR。美洲鲥雌雄个体脑组织转录组中不同类型微卫星的重复基序均具有不同的数量分布特征,其中2核苷酸重复基序数量最多,分别占EST-SSR总数的72.78%和73.60%,而单核苷酸、3核苷酸、4核苷酸、5核苷酸和6核苷酸重复的EST-SSR数量随着重复碱基数量的增加而呈逐级减少趋势;不同EST-SSR重复类型的优势重复基序亦有所不同,单核苷酸重复、2核苷酸重复和3核苷酸重复基序的优势基序分别为A/T、AC/GT和AGG/CCT。从7 671个雌雄美洲鲥脑组织转录组共有EST-SSR中成功鉴定到1 726个多态性EST-SSR,其中鉴定自单核苷酸重复、2核苷酸重复、3核苷酸重复的多态性EST-SSR分别有705,827和116个。20个4核苷酸重复的EST-SSR中,有17个位点多态性较好,其检测1龄美洲鲥的观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)均值分别为0.498,0.620和0.561,表明本单位科研基地的美洲鲥养殖群体历经数年的人工养殖,仍然具有相对较高的遗传多样性。【结论】基于美洲鲥脑组织转录组数据规模化开发了多态性EST-SSR,这些位点可用于美洲鲥养殖群体的遗传多样性评估及分子标记辅助遗传育种。 展开更多
关键词 美洲鲥 高通量测序 脑组织转录组 微卫星标记(SSR) 表达序列标签SSR(est-SSR)
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从中国对虾ESTs中筛选微卫星标记的研究 被引量:38
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作者 徐鹏 周令华 +1 位作者 田丽萍 相建海 《水产学报》 CAS CSCD 北大核心 2003年第3期213-218,共6页
利用生物信息学方法在含有10446个中国对虾ESTs的数据库中进行微卫星序列的筛选,共发现微卫星序列229个,占整个ESTs数据库的2.19%,其中含双碱基重复序列146个和3碱基重复序列58个,分别占在ESTs数据库中发现微卫星序列总数的63.76%和25.3... 利用生物信息学方法在含有10446个中国对虾ESTs的数据库中进行微卫星序列的筛选,共发现微卫星序列229个,占整个ESTs数据库的2.19%,其中含双碱基重复序列146个和3碱基重复序列58个,分别占在ESTs数据库中发现微卫星序列总数的63.76%和25.33%,大部分发现的微卫星序列均为Perfect形式的重复序列。根据筛选得到的微卫星序列设计并合成引物19对进行多态性检测,在有扩增产物的16对引物中,首次筛选得到8个中国对虾微卫星标记,并对这些微卫星标记进行了等位基因频率、观测杂合度、期望杂合度、PIC值等统计学指标的评价。结果表明,在8个微卫星位点上,等位基因的数目从5到15不等,等位基因长度从165~305bp,期望杂合度和多态性信息含量分别为0.59到0.89和0.56到0.88,表明这8个中国对虾微卫星标记完全适合于遗传分析。 展开更多
关键词 中国对虾 estS 微卫星标记 生物信息学 微卫星序列 等位基因频率 观测杂合度 期望杂合度
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刺参(Apostichopus japonicus)EST序列中微卫星分布分析及其标记的筛选 被引量:20
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作者 孙国华 杨建敏 +3 位作者 宋志乐 孙孝德 王卫军 张宇 《海洋与湖沼》 CAS CSCD 北大核心 2010年第1期133-139,共7页
利用微卫星查找软件对现已公布的6632条刺参ESTs的数据库中2—6个碱基重复单元组成的简单序列重复进行筛选,进而对其微卫星的丰度和分布进行比较研究。共发现微卫星序列416个,占整个ESTs数据库的6.48%;其中含双碱基重复序列146个,数量最... 利用微卫星查找软件对现已公布的6632条刺参ESTs的数据库中2—6个碱基重复单元组成的简单序列重复进行筛选,进而对其微卫星的丰度和分布进行比较研究。共发现微卫星序列416个,占整个ESTs数据库的6.48%;其中含双碱基重复序列146个,数量最多,占ESTs数据库中发现微卫星序列总数的65.86%;三、四、五、六碱基重复序列分别占微卫星序列总数的26.68%、1.68%、0.24%、5.53%。对含有SSR位点符合微卫星引物设计的EST序列利用Primer软件结合人工方法设计合成引物21对,其中13对有扩增产物且均为多态位点。扩增产物变性聚丙烯酰胺胶电泳,统计每个微卫星标记等位基因数目,计算等位基因频率、观测杂合度、期望杂合度等统计学指标的评价。结果表明,在13个微卫星位点上,等位基因的数目从3到8不等,等位基因长度从175—382bp,观测杂合度和期望杂合度分别为0.158—0.650和0.198—0.841,所筛选微卫星标记可于遗传分析。 展开更多
关键词 刺参 微卫星 表达序列标签 等位基因
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杨树EST-SSR标记的开发 被引量:36
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作者 张新叶 宋丛文 +2 位作者 张亚东 杨彦伶 黄敏仁 《林业科学》 EI CAS CSCD 北大核心 2009年第9期53-59,共7页
利用来自美洲黑杨及欧美杨的20023条EST序列,进行EST-SSR标记开发研究。首先对这些EST序列进行序列拼接,然后在非重复序列中发现由二核苷酸至五核苷酸组成的SSR重复序列。共在10816个非重复序列中发现1604个序列含有SSR,占总非重复序列... 利用来自美洲黑杨及欧美杨的20023条EST序列,进行EST-SSR标记开发研究。首先对这些EST序列进行序列拼接,然后在非重复序列中发现由二核苷酸至五核苷酸组成的SSR重复序列。共在10816个非重复序列中发现1604个序列含有SSR,占总非重复序列的14.8%。在发现的1918个SSR重复序列中,二核苷酸重复是最丰富的重复类型,占总类型的62.3%。设计合成48对SSR引物,利用6个不同的杨树品种对其进行验证。结果约90%的引物获得扩增产物,58%的引物在6个不同杨树品种中产生多态性分离。另外,还对重复序列的数量及重复类型进行了讨论。 展开更多
关键词 杨树 表达序列标签(est) 简单重复序列(SSR)
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白菜的EST标记及其对油菜的通用性 被引量:20
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作者 忻雅 崔海瑞 +2 位作者 张明龙 林容杓 崔水莲 《遗传》 CAS CSCD 北大核心 2005年第3期410-416,共7页
根据白菜的表达序列标签,设计了28对引物。在对引物、dNTP、MgCl2的浓度及退火温度等参数进行测试后,建立了合适的PCR反应体系。在此反应体系下,以构建EST的白菜自交系A的DNA为模板,对设计的引物进行了筛选,发现有18对引物能对白菜DNA... 根据白菜的表达序列标签,设计了28对引物。在对引物、dNTP、MgCl2的浓度及退火温度等参数进行测试后,建立了合适的PCR反应体系。在此反应体系下,以构建EST的白菜自交系A的DNA为模板,对设计的引物进行了筛选,发现有18对引物能对白菜DNA扩增出产物。用筛选出来的引物分别对17个白菜类品种进行PCR扩增,用琼脂糖凝胶电泳分析其产物的多态性,发现10对引物有多态性,占筛选引物的55.6%。为检测白菜EST标记的通用性,进一步利用设计的引物对不同油菜品种的DNA进行PCR扩增。在检测的28对引物中,共有24对引物能扩增出产物,占引物总数的85.7%,显示多态性的引物为18对,占引物总数的64.3%.。在对白菜DNA能扩增出产物的18对引物中,对油菜完全可用,且有13对引物产生多态性。而在那些对白菜未扩增出产物的10对引物中,也有6对能扩增出产物,其中5对显示多态性。结果证明,通过EST建立分子标记是可行的,而且这种标记对近缘物种是可通用的。 展开更多
关键词 白菜 表达序列标签(est) 分子标记 通用性
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鼻咽癌中染色体3p21区域一个表达下调的EST的鉴定 被引量:14
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作者 贺修胜 陈主初 +6 位作者 田芳 肖志强 贺智敏 关勇军 李峰 何春梅 袁建辉 《癌症》 SCIE CAS CSCD 北大核心 2003年第1期1-5,共5页
背景与目的:研究显示鼻咽癌细胞3p14-25存在高频率杂合性丢失位点。本研究拟寻找与筛选染色体3p21区域与鼻咽癌相关的表达序列标签(expressedsequencetag,EST),为定位候选克隆鼻咽癌相关新基因奠定基础。方法:充分利用网上的生物信息资... 背景与目的:研究显示鼻咽癌细胞3p14-25存在高频率杂合性丢失位点。本研究拟寻找与筛选染色体3p21区域与鼻咽癌相关的表达序列标签(expressedsequencetag,EST),为定位候选克隆鼻咽癌相关新基因奠定基础。方法:充分利用网上的生物信息资源,采用定位查找ESTs,对ESTs进行同源性比较分析、筛选;运用逆转录PCR(reversetranscription-PCR,RT-PCR)方法,检测ESTs在鼻咽癌和正常鼻咽组织中的表达;并用Northernblot杂交方法,检测EST在人其他正常组织及肿瘤细胞系的表达状况。结果:在3p21区域筛选到一个在鼻咽癌中表达下调的EST(N31985),在60.00%(3/5)的鼻咽癌细胞株及47.06%(16/34)的鼻咽癌活检组织检测到有EST(N31985)表达下调,与正常鼻咽上皮组织相比较,差异有统计学意义(P<0.05)。结论:染色体3p21区域EST(N31985)在鼻咽癌中表达下调,提示其可能参与鼻咽癌癌变过程。 展开更多
关键词 基因表达 RT-PCR 鼻咽癌 3p21区域 est
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银杏EST序列中微卫星的分布特征 被引量:30
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作者 樊洪泓 李廷春 +2 位作者 李正鹏 林毅 蔡永萍 《基因组学与应用生物学》 CAS CSCD 北大核心 2009年第5期869-873,共5页
本文利用从NCBI下载的21590条银杏EST序列,分析了银杏(表达序列标签微卫星)EST-SSR在银杏EST序列的分布和比较了在不同长度EST序列中的SSR特性。在剔除冗余和低质量序列后,得到总长为5708.385kb的无冗余EST序列7961条,发现了405个EST... 本文利用从NCBI下载的21590条银杏EST序列,分析了银杏(表达序列标签微卫星)EST-SSR在银杏EST序列的分布和比较了在不同长度EST序列中的SSR特性。在剔除冗余和低质量序列后,得到总长为5708.385kb的无冗余EST序列7961条,发现了405个EST序列(5.09%)含有475个SSR,长度400~1000bp的EST序列含SSR位点数为445个,占SSR总数的93.68%。二核苷酸和三核苷酸基元类型是银杏EST-SSR的主要类型,分别占SSR总数的73.89%和24.00%,最常见的SSR基元是:(AT)n、(AG)n、(AC)n、(AAG)n和(AAT)n。通过对银杏EST序列中SSR位点信息的发掘分析,为有针对性地设计EST-SSR引物,开发银杏EST-SSR分子标记奠定基础。 展开更多
关键词 银杏 表达序列标签 est-SSR 重复基元
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基于EST信息的百合SSR标记的建立 被引量:41
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作者 杨素丽 明军 +2 位作者 刘春 穆鼎 李名扬 《园艺学报》 CAS CSCD 北大核心 2008年第7期1069-1074,共6页
依据已知的百合EST(expres sedsequence tags)序列信息,开发新的SSR(simple sequence re-peats)标记,在NCBI的EST数据库1688条EST中检索到98条含有101个SSR的序列,SSR的检出率为5.98%,其中三核苷酸重复类型占主导地位,出现频率为2.84%。... 依据已知的百合EST(expres sedsequence tags)序列信息,开发新的SSR(simple sequence re-peats)标记,在NCBI的EST数据库1688条EST中检索到98条含有101个SSR的序列,SSR的检出率为5.98%,其中三核苷酸重复类型占主导地位,出现频率为2.84%。EST-SSR的重复基元共搜索到47种,其中三核苷酸重复基元类型最为丰富,大约占重复基元类型总数的一半。利用部分EST-SSRs序列共设计23对SSR引物,以铁炮百合‘Snow Queen’DNA为模板,对引物进行筛选,其中18对引物有扩增产物,占所设计引物总数的78.26%;进一步用这些引物在5个杂种系列13个百合品种进行多态性测试,显示多态性的引物占可扩增引物的66.7%。本研究结果证明了基于百合EST信息建立SSR标记是一种有效而又可行的方法。 展开更多
关键词 百合 est SSR 引物设计
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麦红吸浆虫唾腺EST-SSRs的信息分析及分子标记筛选 被引量:13
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作者 段云 吴仁海 +4 位作者 罗礼智 武予清 蒋月丽 苗进 巩中军 《昆虫学报》 CAS CSCD 北大核心 2011年第10期1147-1154,共8页
昆虫EST资源库的扩充为开发新的分子标记提供了宝贵的资源。本研究对NCBI的EST数据库中来源于麦红吸浆虫Sitodiplosis mosellana唾腺的1217条EST序列进行了unigene组装、SSR信息分析和EST-SSR分子标记筛选。结果表明:在1047个unigenes... 昆虫EST资源库的扩充为开发新的分子标记提供了宝贵的资源。本研究对NCBI的EST数据库中来源于麦红吸浆虫Sitodiplosis mosellana唾腺的1217条EST序列进行了unigene组装、SSR信息分析和EST-SSR分子标记筛选。结果表明:在1047个unigenes中共找到141个SSR位点,分布于106个(10.12%)unigenes中,平均每3.49kb出现一个SSR位点。在1~6碱基重复基元中,1~3碱基是主要重复类型,占总SSR的97.16%以上。A/T(31.21%),AC/GT(15.60%)和AAC/GTT(9.22%)分别是单、双和三碱基中占优势的重复基元类型。利用Primer Premier5.0软件对查找的EST-SSRs进行引物设计,并以麦红吸浆虫基因组DNA为模板,对从中选出的26对SSR引物进行多态性检测。结果有20对(76.92%)引物能扩增出清晰的目的条带,并且其中9对(45%)引物表现出多态性。多态性分析结果表明,从9对EST-SSR引物中,共检测到51个等位基因,平均每个位点含有等位基因数为5.67,平均期望杂合度为0.65,平均多态信息含量为0.60。本研究能够为今后麦红吸浆虫的种群遗传结构与遗传多样性研究提供帮助。 展开更多
关键词 麦红吸浆虫 表达序列标签 est-SSR 分子标记 多态性
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