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《Journal of Fishery Sciences of China》 2011-01
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Construction and analysis of gonad suppression subtractive hybridi-zation library of rice field eel(Monopterus albus)

QU Xiancheng1,SHANG Xiaoli1,CHENG Cui1,QU Xuewei2,ZHANG Yong1,ZHANG Kaiyue1, JIANG Jiaoyun1 1.College of Fisheries and Life Sciences,Shanghai Ocean University,Shanghai 201306,China; 2.Muping District Yantai Fisheries Technical Service Center,Yantai 264100,China  
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 DH5α.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 ran-domly 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 se-quenced 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.
【Fund】: 上海市重点学科建设项目(S30701)
【CateGory Index】: S917.4
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