| 小鼠睾丸特异表达基因TSEG-1的克隆及序列分析 |
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| 引用本文： | 顾朝辉,童强松,曾甫清,刘媛,王智宇,郑丽端,蔡嘉斌,蒋国松.小鼠睾丸特异表达基因TSEG-1的克隆及序列分析[J].遗传,2008,30(3):352-358. |
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| 作者姓名： | 顾朝辉 童强松 曾甫清 刘媛 王智宇 郑丽端 蔡嘉斌 蒋国松 |
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| 作者单位： | 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 |
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| 基金项目： | 国家冉然科学基金，教育部跨世纪优秀人才培养计划 |
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| 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1 668~2 011 kb间克隆出一新基因TSEG-1, 全长为510 bp, 开放阅读框为336 bp, 编码111氨基酸, 分子量12.84258 kDa, 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2a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680 bp。 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。
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| 关 键 词： | 小鼠 睾丸 表达序列标签 TSEG-1 生物信息学 |
| 收稿时间： | 2007-09-15 |
| 修稿时间： | 2007年9月15日 |
Cloning and sequence analysis of TSEG-1, a novel gene specifically expressed in mouse testis |
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| GU Chao-Hui,TONG Qiang-Song,ZENG Fu-Qing,LIU Yuan,WANG Zhi-Yu,ZHENG Li-Duan,CAI Jia-Bin,JIANG Guo-Song.Cloning and sequence analysis of TSEG-1, a novel gene specifically expressed in mouse testis[J].Hereditas,2008,30(3):352-358. |
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| Authors: | GU Chao-Hui TONG Qiang-Song ZENG Fu-Qing LIU Yuan WANG Zhi-Yu ZHENG Li-Duan CAI Jia-Bin JIANG Guo-Song |
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| Affiliation: | Department of Surgery, Huazhong University of Science and Technology, Wuhan 430022, China. zgwhgch@163.com |
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| Abstract: | 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 en-codes 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 demonstrated 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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| Keywords: | 9TSEG-1 |
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