青海湖裸鲤eif5b基因克隆与初步功能分析

Cloning and Preliminary Function Analysis of eif5b Gene in Gymnocypris przewalskii

【目的】围绕青海湖裸鲤卵巢中的eif5b基因克隆和功能开展相关研究,为青海湖裸鲤的卵巢发育机制研究提供理论基础数据。【方法】采用RACE和RT-PCR克隆青海湖裸鲤eif5b全长cDNA序列,对其进行生物信息分析,通过表达研究对其功能进行初步分析。【结果】青海湖裸鲤eif5b全长cDNA序列为2209 bp,包含1293 bp开放阅读框,编码430个氨基酸。氨基酸序列同源性比较发现,克隆的eif5b与其他脊椎动物的亲缘性相近,同源性为75.7%~95.1%,处于系统进化树上同一分支;eif5b与其邻近基因lipt1的共线性在整个脊椎动物中高度保守。【结论】青海湖裸鲤eif5b表达于脑、垂体、鳃、心脏、脾脏、卵巢、精巢和肾脏,在3、6、9、12、18、24月龄卵巢中均存在表达。

【Objective】The related research on cloning and function of eif5b gene in Gymnocypris przewalskii ovary is conducted,which provide the theoretical data for ovary developmental mechanism of G.przewalskii.【Method】The full-length cDNA of eif5b is cloned from G.przewalskii by using RT-PCR and RACE,and the biological information analysis and functional analysis are conducted as well through expression research.【Result】The cloned eif5b gene is 2209 bp,containing a 1293 bp open reading frame which encoded 430 amino acids.The amino acid sequence homology alignment shows that the cloned eif5b is similar to other vertebrates with the similarity between 75.7%—95.1%,which is at a same branch of phylogenetic tree.The colinearity of eif5b and its adjacent gene lipt1 are highly conserved across vertebrates.【Conclusion】The eif5b of G.przewalskii is expressed in brain,pituitary,gill,heart,spleen,ovary,testis,and kidney,and is expressed in ovary with 3-,6-,9-,12-,18-,24-month old.