中华绒螯蟹血细胞cDNA文库的构建及质量初检

为了筛选中华绒螯蟹Eriocheir sinensis血细胞免疫的相关基因,促进蟹类免疫防治的进展,采用Gubler-Hoffman法构建了中华绒螯蟹血细胞的cDNA文库。结果表明：该文库初始滴度为4.50×106cfu/mL,库容为3.3×106cfu/mL,重组率为73%,插入片段大小多为800~2 500 bp。随机挑取了94个重组子进行测序初检,共得到93条平均长度为511 bp的表达序列标签（EST）序列,对所测EST序列进行拼接,得到75条一致性序列,包括15条重叠群（Contigs）和60条单一序列（Singleton）。对测序结果进行比对分析,获得了15条已报道有一定功能的一致性序列,其中4条与免疫相关;另外60条一致性序列还未见报道。研究表明,中华绒螯蟹血细胞cDNA文库已构建成功,且质量较高,从而为开展中华绒螯蟹功能基因克隆、分析和功能基因组学研究奠定了基础。

Establishment of cDNA library from hemocytes of Chinese mitten crab Eriocheir sinensis

A cDNA library for the hemocytes of Chinese mitten crab Eriocheir sinensis was successfully constructed by Gubbler-Hoffman technique and was randomly sequenced.There was 3.3×106cfu/mL in the capacity of library with DNA inserts of 800-2 500 bp,and the recombination rate of 73%.Ninety-three express sequence tags（ESTs） were obtained by gene sequencing of ninety-four recombinants picked up randomly.In addition,75 non-repetitive sequences were formed including 15 contigs and 60 singletons from these ESTs after initial assembly.Fifteen EST sequences were obtained by BLAST analysis,in which whose certain functions were reported while the other 59 sequences were not reported,indicated that the quality of this cDNA library was very good.This cDNA library provided a valuable resource for the functional genome research of the Chinese mitten crab,and will form a fundamental base for further screen and clone of the immune-related genes.