桑树幼叶cDNA文库的构建及部分表达序列标签分析  

Construction and Primary Analysis of cDNA Library From Young Mulberry Leaves

作　　者：方荣俊[1,2] 戚金亮[3] 扈冬青[1,2] 赵卫国[1,2] 汪伟[1,2] 张林[2] 刘利[2] 潘刚[2] 沈兴家[1,2] 潘一乐[2] 杨永华[3] 

机构地区：[1]江苏科技大学生物与环境工程学院,江苏镇江212018 [2]中国农业科学院蚕业研究所,农业部家蚕生物技术重点开放实验室,江苏镇江212018 [3]南京大学生命科学学院,南京210093

出　　处：《蚕业科学》2008年第4期581-586,共6页ACTA SERICOLOGICA SINICA

基　　金：国家科技基础条件平台工作子项目(编号2005DKA21002-09);国家"十一五"科技支撑计划项目(编号2006BAD06-B03);博士后科学基金项目(编号20060400926;0602004-C);公益性行业(农业)科研专项(编号nyhyzx07-020-02)

摘　　要：基于为鉴定和克隆桑树功能基因提供基础信息的目的,采用RNA转录5′末端转换(SMART)法构建了桑树幼叶全长cDNA文库。该文库容量为1.02×106pfu/mL,重组率95%,符合构建基因文库的质量要求。从构建的桑树幼叶cDNA文库中随机挑取48个克隆进行表达序列标签(EST)测序,有效序列为32条,经UniGene数据库归并后为32条,UniGene比率为100%;与NCBI核酸数据库进行比对、查询和注释,在32条序列中有29条序列具有同源性,其中16条为全长序列,完整性比率为55.2%;初步发现具有已知功能基因的ESTs 6个,具有推测功能基因的ESTs 5个,未命名或未知功能基因的ESTs 21个。In order to isolate and clone the functional genes involved in the growth development of mulberry,the full-length cDNA library from young mulberry leaves was constructed using the method of SMART （switching mechanism at 5＇ end of RNA transcript） in this paper. The library consisted of 1.02×10^6 pfu/mL with the recombinant percentage of 95%, which can meet the quality requirement for cDNA library construction. Fortyeight clones were randomly selected from the cDNA library and the ESTs （expressed sequence tags） were sequenced. The 32 effective sequences obtained were merged and the UniGene. ratio reached to 100%. By biasting 32 effective sequences against the NCBI nonredundant nucleotide databases, we found 29 homologous sequences which contained 16 fulllength sequences, and the integrity ratio of the fulllength sequences was 55.2%. Six ESTs with known function, 5 ESTs with putative function and 21 ESTs with unnamed or unknown function were identified, The present results have established a solid molecular base for identifying and cloning functional genes from mulberry in the future.

关 键 词：桑树 CDNA文库 RNA转录5′末端转换 表达序列标签 

分 类 号：S888.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]