| 文蛤肠、外套膜和肝胰脏组织 cDNA 文库构建及其 ESTs 序列分析 |
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| 引用本文： | 高祥刚,李云峰,宋文涛,王健,傅立元,刘卫东,鲍相渤,赫崇波.文蛤肠、外套膜和肝胰脏组织 cDNA 文库构建及其 ESTs 序列分析[J].中国水产科学,2010,17(2). |
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| 作者姓名： | 高祥刚 李云峰 宋文涛 王健 傅立元 刘卫东 鲍相渤 赫崇波 |
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| 作者单位： | 1. 辽宁省海洋水产科学研究院,辽宁省海洋水产分子生物学重点实验室,辽宁,大连,116023
2. 辽宁省海洋水产科学研究院,辽宁省海洋水产分子生物学重点实验室,辽宁,大连,116023;辽宁师范大学,生命科学学院,辽宁,大连,116029 |
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| 基金项目： | 辽宁省科学技术计划重大、重点项目，辽宁省海洋与渔业科研计划项目，国家海洋公益性行业科技专项 |
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| 摘 要： | 利用SMART cDNA文库构建试剂盒构建了文蛤(Meretrix meretrix)肠、外套膜和肝胰脏组织的cDNA文库。经测定原始文库滴度分别为2.10×106、1.70×106和1.60×106,重组率高于95%,肠组织文库插入片段长度均大于1000bp,外套膜和肝胰脏组织文库的插入片段长度大于1kb的占87.5%,文库质量符合标准要求。随机选取文库的3168个克隆进行5′端测序,获得高质量表达序列标签(Expressed Sequence Tags,ESTs)3029个(肠:1005,外套膜:1019,肝胰脏:1005),测序成功率95.58%。经质量控制和拼接得到1796个单基因簇(Unigene),其中306个叠联群(Contigs),1490个单一序列(Singletons)。通过Blastx搜索比对、查询和注释分析,共得到已知基因696个(肠:216,外套膜:235个;肝胰脏:245个),占总数38.75%。在1796个单基因簇(Unigene)发现微卫星序列55条,这些存在微卫星位点的序列占整个ESTs数据库的3.1%。
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| 关 键 词： | 文蛤 cDNA文库 表达序列标签 微卫星标记 |
| 修稿时间： | 2010/3/30 0:00:00 |
Construction of cDNA libraries from intestine,mantle and hepatopancreas of hard clam (Meretrix meretrix) and ESTs analysis |
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| GAO Xianggang,LI Yunfeng,SONG Wentao,WANG Jian,FU Liyuan,LIU Weidong,BAO Xiangbo,HE Chongbo.Construction of cDNA libraries from intestine,mantle and hepatopancreas of hard clam (Meretrix meretrix) and ESTs analysis[J].Journal of Fishery Sciences of China,2010,17(2). |
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| Authors: | GAO Xianggang LI Yunfeng SONG Wentao WANG Jian FU Liyuan LIU Weidong BAO Xiangbo HE Chongbo |
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| Institution: | GAO Xianggang 1,LI Yunfeng 1,SONG Wentao 1,2,WANG Jian 1,FU Liyuan 1,LIU Weidong 1,BAO Xiangbo 1,HE Chongbo 1 (1.Liaoning Ocean , Fishery Science Institute,Liaoning Key Laboratory of Marine Fishery Molecular Biology,Dalian 116023,China,2.College of Life Sciences,Liaoning Normal University,Dalian 116029,China) |
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| Abstract: | The cDNA libraries of intestine, mantle and hepatopancreas of hard clam (Meretrix meretrix)were constructed with SMART cDNA Library Construction Kit. The libraries have a high titer of 2.10×10~6, 1.70×10~6 and 1.60×10~6, respectively. The recombined efficiency of each cDNA library exceeded 95%. The all insert sizes of intestine library were over 1 kb, and the insert sizes over 1 kb in the mantle and hepatopancreas libraries were over 87.5%. Atotal of 3 168 clones of the libraries were randomly picked and sequenced from the 5' end of the cDNAs using a M13 universal primer, and 3 029 raw sequences of the ESTs were processed and then assembled into 1 796 unigene involving 306 contigs and 1 490 singletons. Blastx analysis showed that 696 unigenes (216 for intestine, 235 for mantle and 245 for hepatopancreas) (38.75%) had significant homology (Evalue ≤10~(-5)) to genes with known or putative functions in GenBank.The sequences containing microsatellite DNA (SSR) accounted for 3.1% of all ESTs indicating that SSR sequences were rich in the ESTs of hard clam. |
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| Keywords: | Meretrix meretrix cDNA Library expressed sequence tags EST-SSR |
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