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小鼠睾丸特异表达基因TSEG-1的克隆及序列分析
  • ISSN号:0253-9772
  • 期刊名称:《遗传》
  • 时间:0
  • 分类:Q954.434[生物学—动物学] S512.1[农业科学—作物学]
  • 作者机构:[1]华中科技大学同济医学院附属协和医院外科,武汉430022, [2]华中科技大学同济医学院附属协和医院病理科,武汉430022
  • 相关基金:国家自然科学基金资助项目(编号:30200284,30600278,30772359)和教育部新世纪优秀人才支持计划
中文摘要:

从表达序列标签(expressed sequence tags,ESTs)数据库ZooDDD中获得小鼠正常睾丸表达的EST,通过dbEST数据库检索出与其高度同源的EST序列,构建EST叠加群(contigs),Biolign软件拼接,GeneScan软件预测contigs对应的基因组序列中的外显子、内含子 针对开放阅读框设计引物序列,采用RT-PCR从小鼠睾丸组织中克隆新基因的cDNA,分析该基因在小鼠各脏器中的mRNA表达,并对测序结果进行生物信息学分析。结果表明:在小鼠X染色体的1668~2011kb间克隆出一新基因TSEG-1,全长为510bp,开放阅读框为336bp,编码111氨基酸,分子量12.84258kDa,等电点11.4000。RT-PCR证实该基因开放阅读框正确,在小鼠睾丸组织中特异性表达,且与小鼠其他cDNA无同源性,获得GenBank登录号EU079024。功能区分析发现TSEG-1蛋白可能为一种跨膜蛋白,跨膜区位于第41~61氨基酸残基。TSEG-1基因与人类睾丸特异性组蛋白2α变异体基因有较高同源性,在TSEG-1基因5′-端非编码侧翼预测发现存在1个启动子区域,范围为680bp。TSEG-1蛋白可能有4个抗原性位点,2个特异性蛋白激酶的磷酸化位点,其亚细胞定位可能位于线粒体。小鼠睾丸特异性基因TSEG-1的克隆为进一步研究其生物学功能和表达调控奠定了基础。

英文摘要:

The expressed sequence tags (ESTs) of normal mouse testis were obtained from online EST database ZooDDD. Their highly homologous EST sequences were found through the dbEST database to construct contigs, and spliced by the biomedical software Biolign. The corresponding exons and introns within genome sequences were predicted by software GeneScan. According to the open reading frame, the primers were designed. RT-PCR was applied in the cloning of novel gene from mouse testis and analyzing its expression pattern in various mouse tissues. The bioinformatics analysis on the sequencing results of TSEG-1 was conducted. Results indicated that a novel gene TSEG-1 was cloned from 1 668-2 011 kb of mouse X chromosome, with full-length sequence of 510 bp. The open reading frame (ORF) is 336 bp in length and encodes a deduced amino acid sequence of 111 residues. The molecular weight of TSEG-1 protein is 12.84258 kDa, and its pI is 11.4000. RT-PCR demonstlated the correctness of its ORF. TSEG-1 was distinctively expressed in testis, but not in other tissues of mouse. No obvious homology with other mouse cDNA was found for TSEG-1. The GenBank accession number EU079024 was achieved. It was predicted that TSEG-1 is a kind of transmembrane protein, and the transmembrane domain is from 41 amino acid residue to 61 amino acid residue. Blastn analysis revealed its high homology to human testis-specific gene H2AX. Computational prediction of the 5′-untranslated region of TSEG-1 gene revealed a 680 bp-length promoter region. There are four antigen binding sites and two phosphorylation sites of specific protein kinase in TSEG-1 protein, with subcellular localization in mitochondria. The cloning of mouse testis specific gene TSEG-1 laid a foundation for subsequent research of its biological function and expression regulation.

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期刊信息
  • 《遗传》
  • 中国科技核心期刊
  • 主管单位:中国科学院
  • 主办单位:中国遗传学会
  • 主编:张永清
  • 地址:北京朝阳区北辰西路1号院中国科学院遗传发育所
  • 邮编:100101
  • 邮箱:yczz@genetics.ac.cn
  • 电话:010-64807669
  • 国际标准刊号:ISSN:0253-9772
  • 国内统一刊号:ISSN:11-1913/R
  • 邮发代号:2-810
  • 获奖情况:
  • 中国自然科学核心期刊,《CAJ-CD》执行优秀奖,2008年12月获“中国精品科技期刊”证书和北京市印...
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  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,荷兰文摘与引文数据库,美国生物医学检索系统,美国生物科学数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:23270