家蚕黄茧近等基因系SSH文库的构建及部分EST序列分析  

SSH cDNA Library Construction and Corresponding EST Analysis of NILs for Yellow Cocoon Gene in Silkworm,Bombyx mori

作　　者：李刚[1] 徐安英[2] 李木旺[2] 陈兆羽[1] 张月华[2] 钱荷英[2] 孙平江[2] 郭锡杰[1] 

机构地区：[1]江苏科技大学,江苏镇江212003 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2009年第3期486-493,共8页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB12-1004);江苏省科技项目(编号BG2007322)

摘　　要：以家蚕黄茧品系KY(YYCC)和白茧品系C108(+Y+Y+C+C)组配黄茧(C)近等基因系。挑取回交11代的黄茧(C)近等基因系群体中的黄血个体的黄色和浅黄色中部丝腺作为实验材料,构建家蚕黄茧近等基因系抑制消减杂交(SSH)cDNA文库。通过对SSHcDNA文库中随机挑选的104个阳性克隆测序和聚类拼接,得到15个假定基因(contig)、25个已知功能基因(unigenes)和5个未知功能基因(unknown)。获得的表达序列标签(EST)经Blast比对和同源性分析,初步发现这些基因参与家蚕体内能量代谢、基因转录、信号传导、物质转运、细胞生长分裂、细胞结构形成、蛋白质合成与降解、核酸及蛋白质加工、机体防御等诸多生物学过程。分析结果可作为深入研究家蚕茧着色分子机制的基础信息。Silkworm near-isogenic lines（NILs） of yellow cocoon（C） gene were constructed using silkworm yellow cocoon strain KY（YYCC） and white cocoon strain C108（＋Y＋Y＋C＋C）.Individuals with yellow and yellowish middle silk glands were selected from yellow hemolymph individuals among the yellow cocoon NIL population of the 11th backcross generation to construct the subtracted cDNA library using PCR-SelectTM cDNA Subtraction Kit（Clontech）.Plasmids were extracted from 104 positive clones randomly selected from the library and then sequenced and assembled using CAP3 software.As a result,16 deduced genes（contigs）,25 unigenes and 5 unknown genes were obtained.BLASTx searches and homology analysis with existing expressed sequence tags（ESTs） revealed that these genes were involved in many biological processes such as energy metabolism,gene transcription,signal transduction,substance transportation,cell growth and division,protein synthesis or degradation,nucleic acid and protein processing,and organismal defense etc.Our results provide useful fundamental information for further studies on molecular mechanism of color formation in silkworm cocoons.

关 键 词：家蚕 黄茧基因 抑制性消减杂交 近等基因系 表达序列标签 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]