中华鲟piwil1基因的克隆表达特征研究  

Molecular cloning and expression analysis of piwil1 gene in Acipenser sinensis

作　　者：李创举[1,2] 杨晓鸽[1] 岳华梅[1,2] 叶欢[1] 危起伟[1,2] 

机构地区：[1]农业部淡水生物多样性保护重点实验室,中国水产科学研究院长江水产研究所,武汉430223 [2]中国水产科学研究院淡水渔业研究中心,江苏无锡214081

出　　处：《淡水渔业》2016年第6期20-25,共6页Freshwater Fisheries

基　　金：中国水产科学研究院淡水渔业研究中心公益性科研院所基本科研业务费专项资金项目(2015JBFM41);公益性行业(农业)科研专项(201203086);国家自然科学基金项目(31472286)

摘　　要：Piwi是鱼类配子发生和性腺发育的重要调控因子。本研究从中华鲟(Acipenser sinensis)性腺中克隆得到了piwi基因的全长c DNA序列,该序列长3 393 bp,开放阅读框为2 583 bp,编码860个氨基酸。氨基酸序列多重对比发现,该序列具有PAZ和PIWI保守结构域,与其他鱼类piwil具有较高同源性;系统进化分析显示,中华鲟piwil聚于piwil1分支;因此,所得序列为piwi-like 1基因,命名为Aspiwil1。荧光定量PCR研究表明,Aspiwil1为母源表达,且其表达量随着胚胎发育逐渐降低;Aspiwil1在性腺中大量表达,在脑组织中也有较低水平的表达。性腺组织切片RNA原位杂交结果表明,Aspiwil1仅在生殖细胞表达,且其杂交信号随着卵子发生逐渐增强、精子发生逐渐减弱。本研究为中华鲟配子发生过程中Aspiwil1的功能研究提供了基础。Piwi genes play important roles in regulating the proliferation and division of germline stem cells. A piwi gene,piwi- like 1（ Aspiwil1）,was cloned through homologous cloning method from the gonad of Acipenser sinensis. The full length of Aspiwil1 was 3 393 bp with an opening reading frame（ ORF） of 2 583 bp. Multiple amino acid sequences alignment showed sturgeon As piwil1 contained the typical conserved PAZ and PIWI motif. Phylogenetic analysis revealed that sturgeon Aspiwil1 was clustered into the Piwil1 clade. Aspiwil1 transcripts were found to originate from the maternal parent,of which expression level was gradually decreased with the embryo development by qRT- PCR analyses. In different tissues,Aspiwil1 was distributed in brain besides the gonads. In situ hybridization in gonadal tissues presented that Aspiwil1 transcripts were only detected in germ cells of both sexes. This work would be valuable for further investigation of piwi gene function in the gonadal development of sturgeon.

关 键 词：中华鲟(Acipenser sinensis) PIWI 克隆 表达 原位杂交 

分 类 号：S917.0[农业科学—水产科学]