团头鲂SPATA4基因的分子克隆及表达分析  

Molecular cloning and expression analysis of Megalobrama amblycephala SPATA4

作　　者：韦新兰[1] 张杰[1] 陈丽萍[1] 王卫民[1] 高泽霞[1] 王焕岭[1] 

机构地区：[1]华中农业大学水产学院/农业动物遗传育种与繁殖教育部重点实验室,武汉430070

出　　处：《华中农业大学学报》2013年第3期99-104,共6页Journal of Huazhong Agricultural University

基　　金：教育部新世纪优秀人才计划项目(NCET-10-0404);中央高校基本科研业务专项(2010PY004)

摘　　要：采用RACE(rapid amplification of cDNA ends)技术从团头鲂精巢中克隆出精子发生相关基因4(sper-matogenesis associated gene 4,SPATA4)。该基因cDNA全长为1 076 bp,包括1个678 bp的完整阅读框架,编码225个氨基酸,预测的氨基酸序列的分子质量为25.72 ku,等电点为9.32。序列分析显示团头鲂SPATA4基因与其他脊椎动物同源性较高。荧光定量PCR结果显示SPATA4基因在受精后1～206 h期间,该基因在受精后4 h和受精后28 h的表达量最高,在受精后50 h表达量最低,而在其他时期的表达水平基本一致;在所检测的10个组织中,SPATA4基因在精巢中的表达量最高。The spermatogenesis associated 4 （SPATA4） gene from testis of Wuchang bream （Meg- alobrarna amblycephala） were cloned using RACE （rapid amplification of cDNA ends） technology and characterized. The full length cDNA of SPATA4 was 1 076 bp including a 678 bp open reading frame, which encode a putative protein of 225 amino acids residues with a theoretical molecular weight of 25.72 ku and an isoelectric point of 9.32. Further analysis of SPATA4 sequence indicated that it had high i- dentity with other vertebrates. Quantitative real time PCR was performed to detect the SPATA4 mR- NA in different tissues of adult fish and various developmental stages. The results showed that during 1- 206 h post-fertilization, the SPATA4 gene had the highest expression levels at 4 h and 28 h, decreased at 50 h to the lowest level, and had similar expression levels at other time points. In the ten tissues tested, the SPATA4 gene showed highest expression level in testis.

关 键 词：团头鲂 精巢 SPATA4 克隆 时空表达 

分 类 号：Q959.468[生物学—动物学]