小鼠睾丸新基因TSEG4的克隆及在睾丸中的表达.docVIP

小鼠睾丸新基因TSEG-4的克隆及在睾丸中的表达 1 胡涛1 ，王智宇1，童强松2，曾甫清1，陈晓春1，顾朝辉1，郑丽端3 （430022 武汉,华中科技大学同济医学院附属协和医院：泌尿外科1，小儿外科2，病理科3） [摘要] 目的： 克隆小鼠睾丸特异性基因并检测其表达特征。方 法：从二级表达序列标签（expressed sequence tags, EST）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Biolign软件拼接GeneScan软件预测，采用RT-PCR从小鼠睾丸组织中克隆新基因的cDNA，并对测序结果进行生物信息学分析。real time-PCR法检测其在手术隐睾模型，睾丸扭转模型和小鼠中的表达bp，编码136个氨基酸。GenBank 登录号EU477371。功能区分析发现，TSEG-4编码蛋白有3个酪蛋白激酶(CK2)磷酸化位点2个蛋白激酶C (PKC)磷酸化位点。TSEG-4曲细精管21天开始高表达，直至生后第2个月，而在6月龄小鼠睾丸组织 [关键词] 克隆；睾丸；表达序列标签；生物信息学 ；精子发生 [中图法分类号] [文献标识码] A 基金项目：国家自然科学基金 、教育部新世纪优秀人才支持计划NCET-06-0641)、教育部留学回国人员科研启动基金（2008889） *通作者：童强松，Tel: 86-27 E-mail: qs_tong@ Cloning of a novel mouse testis-specific gene TSEG-4 and its expression in developmental stages of testes Hu Tao 1, Wang Zhiyu 1, Tong QiangSong2, Zeng Fuqing1, Chen Xiaochun1, Gu Zhaohui 1, ZHENG LiDuan 3. （1 Department of Urology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Hubei 430022, China，2 Department of Pediatric Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Hubei 430022, China，3 Department of Pathology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Hubei 430022, China） 【Abstract】 Objective To clone mouse testis-specific gene and analyze its expression pattern. Methods: The expressed sequence tags (EST) expressed in normal mouse testis were obtained from the online EST database ZooDDD. The full-length gene was predicted by using dbEST database, biomedical software Biolign and software GeneScan. RT-PCR was applied in the cloning of novel gene from mouse testis tissue. The sequencing results were resort to bioinformatics analysis. In situ hybridization and real-time PCR were used to measure its expression localization and expression levels in developmental stage of mouse testis. Results: The novel mouse testis gene TSEG-4 was cloned, which locates at chromosome 11qA4. T