咖啡鲆1-6体节cDNA文库的建立及EST分析

Construction and EST Analysis of cDNA Library of the Rosy Barb's 1-6 Somite Develomental Stage

摘要:

本研究采用SMARTTM cDNA文库构建试剂盒构建了咖啡鲆胚胎发育期1-6体节 cDNA文库。经检验，文库的滴度为1.4×106 pfu/mL，阳性克隆率大于80%。对112个随机挑选的克隆进行两端测序，结果显示有98个测序成功。经在GenBank 上BLAST 比对，98个克隆属于83个基因，其中73个为已知功能基因，另外10个没有明显的同源序列。在已知功能的73个基因中，包括2个与骨骼和运动有关的基因、6个与信号传导有关的基因、5个与能量转换有关的基因、8个与蛋白合成相关的基因、3个与生长发育相关的基因、2个与免疫蛋白相关的基因以及27个假想蛋白和20个其他的基因。咖啡鲆1-6体节cDNA文库的建立旨在为咖啡鲆研究奠定基础。

Abstract:

Abstract In the present study a cDNA library of the rosy barb 1-6 somite stage was constructed. The titer of the cDNA library was estimated as~1.4×106 pfu/mL with the recombination rate above 80%. 112 clones with insert≥1kb to sequence were randomly picked and 83 independent sequences were got. BLASTX analysis of the 83 cDNAs showed that the amino acid sequences of the polypeptides deduced from 73 of the clones were strong matches(P<10-10) to those of previously identified proteins, while the other (10 cDNAs) showed no similarity using the present sequence determination and database analysis. On the basis of these results, 2 cDNAs for genes were identified being associated with Cytoskeleton and motility and 6 genes for proteins were related to Signal transduction. And 5 genes were for energy conversion and 8 genes were for protein sythesis. In addition, 3 gens were for development related proteins, and 2 genes were forimmunal proteins. And 27 genes were for hypothetical protein and 20 genes were for others. This study provided a preliminary view of the gene information involved in the development of the rosy barb 1-6 somite stage.