家蚕单性生殖早期胚胎SSH文库的构建及其序列分析  

SSH cDNA Library Construction and Sequence Analysis From the Autogenesis Early Embryo in the Silkworm,Bombyx mori

作　　者：王艳敏[1] 夏定国[2] 张业顺[2] 任晓俊[1] 韦亚东[2] 张国政[1,3] 

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蚕桑遗传改良重点开放实验室,江苏镇江212018 [3]中国农业科学院蚕业研究所,农业部蚕桑遗传改良重点开放实验室,江苏镇江212018

出　　处：《蚕业科学》2009年第3期476-485,共10页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划“863”项目(编号2006AA10A-119);江苏省普通高校研究生科研创新计划项目(编号CX08S-024Z)

摘　　要：为了探讨家蚕早期胚胎性别决定的分子机制,以非减数分裂孤雌生殖（AMP）和双精雄核发育（BSA）技术获得的家蚕单一性别早期（2-24 h）发育蚕卵为材料,利用抑制消减杂交（SSH）技术构建了正向和反向抑制消减杂交cDNA文库。在正向与反向抑制消减杂交文库中分别随机选择62个和46个克隆测序,分别获得43种和29种cDNA序列,其中25个为新的cDNA序列。生物信息学分析显示,抑制消减杂交文库中与单性生殖有关的高温、低温、辐射等诱导表达基因出现的频次较高。对部分文库序列进行半定量RT-PCR分析表明,这些基因在两种单性生殖早期发育胚胎中存在差异表达。In order to explore the molecular mechanism of sex determination of the silkworm early embryos,the embryonic development stage eggs（224 h） of unitary sex of female or male were prepared by artificial autogenesis by means of ameiotic parthenogenesis（AMP） and bispermic androgenesis（BSA） in the silkworm,Bombyx mori.The suppression subtracted hybridization（SSH） cDNA libraries of forward and reverse directions were constructed between the both.62 and 46 clones were selected randomly for sequencing from the forward（female） and reverse（male） subtracted libraries.43 and 29 cDNA sequences were obtained,among which 25 were novel cDNA sequences.Sequence analyses revealed that the genes related to treatments of high temperature or low temperature and radiation etc in autogenesis emerged in a relatively high frequency.The results of semi-quantitative RT-PCR showed that the cDNAs of subtracted library have differential expression in the 024 h artificial autogenesis eggs of unitary sex of female or male.

关 键 词：家蚕 非减数分裂孤雌生殖 双精雄核发育 抑制消减杂交 

分 类 号：S882.4[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]