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悦目金蛛丝腺SMART RACE cDNA文库的构建与鉴定
引用本文:潘鸿春,宋大祥,周开亚.悦目金蛛丝腺SMART RACE cDNA文库的构建与鉴定[J].蛛形学报,2005,14(2):65-69.
作者姓名:潘鸿春  宋大祥  周开亚
作者单位:1. 南京师范大学生命科学学院遗传资源研究所,南京,210097;安徽师范大学生命科学学院,芜湖,241000
2. 南京师范大学生命科学学院遗传资源研究所,南京,210097;河北大学生命科学学院,保定,071002
3. 南京师范大学生命科学学院遗传资源研究所,南京,210097
基金项目:国家高技术研究发展计划(863计划),安徽省教育厅自然科学基金,安徽师范大学校科研和教改项目
摘    要:运用SMART 技术构建了悦目金蛛丝腺SMART RACE cDNA文库.经检测,文库所含全长cDNA的长度主要集中在500 bp~2000 bp 之间;把双链cDNA通过T/A克隆、随机挑选阳性克隆并测序后,得到1条752 bp的全长cDNA序列.以该文库为模板,用依据这条全长cDNA序列设计的基因特异性引物与接头引物进行RACE, 3'RACE 和5'RACE的产物拼接后的全长序列与上述全长cDNA序列一致.结果表明,该文库适于用RACE方法从中分离在悦目金蛛丝腺中表达基因的全长cDNA.本文还对SMART 技术的特点和局限性进行了讨论.

关 键 词:悦目金蛛  丝腺  SMART技术  cDNA文库
收稿时间:2005-08-26
修稿时间:2005年8月26日

Construction and Characterization of SMART RACE cDNA Library of Silk Gland in Argiope amoena
PAN Hong-Chun,SONG Da-Xiang,ZHOU Kai-Ya.Construction and Characterization of SMART RACE cDNA Library of Silk Gland in Argiope amoena[J].Acta Arachnologica Sinica,2005,14(2):65-69.
Authors:PAN Hong-Chun  SONG Da-Xiang  ZHOU Kai-Ya
Abstract:A SMART RACE eDNA library of silk gland in Argiope amoena was constructed successfully with SMART technique. The result showed that the length of the full-length eDNAs in the library was pooled mainly between 500 and 2 000 base pairs. RACE reactions were done with the adaptor primers and the gene-speeifie primers designed according to one EST sequence from silk grand in Argiope amoena. By cloning and sequencing of amplified products, the full-length eDNA sequence of the gene which the EST belongs to was obtained. The library was appropriate for cloning the full-length cDNAs of these genes expressed in the silk gland of Argiope amoena. In addition, the features and limitations of SMART technique were discussed.
Keywords:RACE
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