中华鳖（Pelodiscussinensis）肝脏cDNA文库的构建和EST序列的初步分析

摘要

A high quality cDNA library from the liver of Chinese softshell turtle Pelodiscus sinensis was constructed by Superscript cDNA recombinant technique, and expressed sequence tags (ESTs) were analyzed by Gene Ontology method. The results indicate that the titre of the constructed library was 2.982× 10^6 CFU/ml, the percentage of recombination was 93%, and the average length of inserted cDNA fragment was about lkb. 4146 raw sequences of the ESTs were obtained, 1957 unigenes including 404 contigs and 1553 singletons were found. 264 cDNA sequences showed similarity to known breeding-relevant genes, including ubiquitin C variant 2, heat shock protein70, metallothionein, superoxidedismutase, CD9, antibacterial peptides and so on. These results built the foundation for further researches on germplasm resources evaluation, molecular genetics studies and molecular breeding ofP sinensis.%采用SuperscriptcDNA全长文库构建法构建了高质量的中华鳖肝脏cDNA文库，GO法对ESTs进行注释。结果表明，文库容量为2．982×10^6。CFU／ml，重组率93％，平均插入片段约为1kb。测序获得了4146个有效序列，发现1957条单基因簇，包括404个序列重叠群和1553条单一序列。279个基因簇得到基因注释，发现了多种与育种价值相关的基因，如泛素、热休克蛋白等抗逆基因、金属硫蛋白等抗金属基因、超氧化物歧化酶、CD9等免疫相关基因，为中华鳖分子遗传学研究、种质资源评价和分子选育等奠定了基础。