鲤脑组织cDNA文库的构建及鉴定  

Construction and identification of cDNA library for brain of common carp Cyprinus carpio

作　　者：何薇[1,2] 李言[1,2] 丁雷[1,2] 李勇[1,2] 梁利群[1] 

机构地区：[1]中国水产科学研究院黑龙江水产研究所,黑龙江哈尔滨150070 [2]大连水产学院生命科学与技术学院,辽宁大连116023

出　　处：《大连水产学院学报》2009年第3期195-199,共5页Journal of Dalian Fisheries University

基　　金：国家重点基础研究发展计划973项目(2004CB117405)

摘　　要：用Gubler-Hoffman法构建了耐低温鲤Cyprinus carpio脑组织全长cDNA文库,经测定库容量为5.7×105cfu/mL,文库的重组率接近95%,平均插入片段长度为1 500 bp,符合文库保存和筛选实验的要求。同时,根据鲤与低温相关的消减cDNA文库中低温下特异表达基因的片段序列,设计了PCR引物,并在此cDNA文库中进行PCR检测和序列测定。使用BLAST软件将测序的10个cDNA序列同GenBank等数据库进行比对,结果显示,8个阳性克隆有相关同源性,2个克隆为功能未知的基因,全长率接近75%。Encephalic cDNA library of common carp Cyprinus carpio was constructed by Gubler -Hoffman technique. The capacity of library reached 5.7 ×10^5 cfu/mL with recombinant clones of 95% and the inserts had approximately 1 500 bp in average size. The library was found to meet the demand for both library conservation and filtration. Meanwhile, the primer to set out in the library was designed, and 10 positive clones were sequenced and aligned on Internet, showing higher identities among 8 clones and 2 clones maybe unknown genes and the complete eDNA sequences of 75%. These results indicated that the quality of the encephalic cDNA library about common carp was excellent and provided a useful resource for the functional genomic research related to microthermal traits in common carp.

关 键 词：鲤 脑 CDNA文库 低温性状 

分 类 号：Q959[生物学—动物学]