团头鲂SPATA4基因的分子克隆及表达分析
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教育部新世纪优秀人才计划项目(NCET-10-0404)和中央高校基本科研业务专项(2010PY004)


Molecular cloning and expression analysis of Megalobrama amblycephala SPATA4
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    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(spermatogenesis 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。

    Abstract:

    The spermatogenesis associated 4 (SPATA4) gene from testis of Wuchang bream (Megalobrama 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 identity with other vertebrates. Quantitative real time PCR was performed to detect the SPATA4 mRNA 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.

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韦新兰,张杰,陈丽萍,王卫民,高泽霞,王焕岭.团头鲂SPATA4基因的分子克隆及表达分析[J].华中农业大学学报,2013,32(3):99-104

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  • 收稿日期:2012-06-11
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