中国水产科学2011，Vol.18Issue(1)：23-28,6.DOI:10.3724/SP.J.1118.2011.00023
黄鳝性腺抑制差减文库的构建和分析
Construction and analysis of gonad suppression subtractive hybridization library of rice field eel(Monopterus albus)
摘要
Abstract
By using suppression subtractive hybridization (SSH), subtracted cDNA libraries were constructed from rice field eel (Monopterus albus) gonad at Ⅳ ovary and ovotestis stage. Two-directional (forward and backward) SSH was performed under cDNAs of rice field eel gonad at Ⅳ ovary and ovotestis stage. The cDNA fragments were inserted into plasmid vectors, and then transformed into Escherichia coli DHSα. Finally, forward and backward subtracted cDNA libraries (461 and 674 clones, respectively) were obtained. By PCR detection, length of the subtractive cDNA fragments cloned into the vector ranged from 200 bp to 2 000 bp. One hundred positive clones were selected and sequenced randomly from the forward and the backward libraries, and ninety gene fragment sequences were obtained. EST analysis of these sequences was performed with Blastx and Blaxtn by comparing sequences with GenBank database. Two sequenced genes were further analyzed by semi-quantitative RT-PCR method. The results showed that the library can enrich differentially expressed genes in the two phases gonad.关键词
黄鳝/性腺/抑制性差减杂交/差减cDNA文库分类
农业科技引用本文复制引用
曲宪成,尚晓莉,程翠,曲学伟,张勇,张开岳,蒋骄云..黄鳝性腺抑制差减文库的构建和分析[J].中国水产科学,2011,18(1):23-28,6.基金项目
上海市重点学科建设项目(S30701). （S30701）
