引用本文:	秦玉明,苏秀榕,李晔,马斌,李惠,王孟前,贺静静,李太武.缢蛏(Sinonovacula constricta)cDNA 文库的构建及肌动蛋白基因的研究.海洋与湖沼,2010,41(1):54-60.

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*缢蛏(Sinonovacula constricta)cDNA 文库的构建及肌动蛋白基因的研究*
秦玉明¹, 苏秀榕¹, 李晔¹, 马斌¹, 李惠¹, 王孟前¹, 贺静静¹, 李太武²
1.宁波大学生命科学与生物工程学院;2.宁波大学生命科学与生物工程学院,宁波城市职业技术学院

摘要:

采用SMART(switching metchanism at 5’ end of RNA transcript)技术构建缢蛏cDNA 基因文库。并对cDNA 文库的滴度, 重组率和插入片段的大小进行了检测。结果表明, 该cDNA 文库的滴度为5.50×10⁴ cfu/ml, 重组率为92.5%, 插入片段的长度大多在1kb 以上。对478 个克隆子进行了测序, 共得到430 个有效序列, 其中166 个序列无同源性, 264 条序列具有同源性。从中得到肌动蛋白(actin)cDNA 的全长1538bp, 开放阅读框(ORF)为1131bp, 编码376 个氨基酸, 其预测蛋白的分子量为41.78kD, 等电点为5.30。该氨基酸序列与其它物种的同源性大都在90%以上。

关键词: 缢蛏, cDNA 文库, 序列分析, 肌动蛋白基因

DOI：10.11693/hyhz201001007007

分类号:

基金项目:国家高技术研究发展计划(863 计划), 2006AA10A410 号; 长江学者和创新团队发展计划, 2007—2009; 宁波市科技局资助项目, 2006C100041 号; 宁波市海洋渔业局资助项目, 2006—2008。

附件

CONSTRUCTION OF cDNA LIBRARY WITH SINONOVACULA CONSTRICAT AND SEQUENCE ANALYSIS OF ACTIN GENE

QIN Yu-Ming¹, SU Xiu-Rong¹, LI Ye¹, MA Bin¹, LI Hui¹, WANG Meng-Qian¹, HE Jing-Jing¹, LI Tai-Wu^1,2

1.Faculty of Life Science and Biotechnology, Ningbo University;2.Ningbo City College of Vocational Technology

Abstract:

SMART (switching mechanism at 5'-end of RNA transcript) technique to construct full-length cDNA library with Sinonovacula constricta was adopted and the library titre, recombinant rate, and the length of inserted cDNA were measured. The results indicate that this library’ titre reached 5.50×10⁴ cfu/ml; the percentage of recombination was as high as 92.5%. The PCR results show that the average size of inserts was larger than 1000bp. 478 clones were sequenced and 430 valid sequences were obtained, in which 264 cDNA sequences showed similarity to known genes. Actin was found by aligning it with those of the homologues in the NCBI database’s Blast program. Its full-length was 1538bp, including an open reading frame (ORF) encoding apolyeptide of 376 amino acids with predicted molecular weight of 41.78kD and theoretical isoelectric point of 5.30. The amino acids sequence aligned with other species was higher than 90%, meaning actin is a high conserved gene.

Key words: Sinonovacula constricta, cDNA library, Sequence analysis, Actin gene