缢蛏(Sinonovacula constricta)cDNA文库的构建及肌动蛋白基因的研究  

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

作　　者：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[1,2] 

机构地区：[1]宁波大学生命科学与生物工程学院,宁波315211 [2]宁波城市职业技术学院,宁波315100

出　　处：《海洋与湖沼》2010年第1期54-60,共7页Oceanologia Et Limnologia Sinica

基　　金：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

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5.50×104cfu/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。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 5.50×10^4 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.

关 键 词：缢蛏 CDNA文库 序列分析 肌动蛋白基因 

分 类 号：S968.3[农业科学—水产养殖]