Wang Zhi-Yu, Tong Qiang-Song, Zeng Fu-Qing, Liu Yuan, Gu Zhao-Hui, Zheng Li-Duan, Cai Jia-Bin, Jiang Guo-Song
Department of Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, China.
Zhonghua Nan Ke Xue. 2009 Feb;15(2):99-105.
To clone the mouse testis specific gene TSEG-2 via a bioinformatic approach.
The expressed sequence tags (EST) in the normal mouse testis were obtained from the online EST database ZooDDD. Their highly homologous EST sequences were retrieved through the dbEST database to construct contigs and spliced with the biomedical software Biolign. The corresponding exons and introns within the genome sequences were predicted with the software GeneScan. Primers were designed according to the open reading frame. RT-PCR was applied in cloning the cDNA of the novel gene from the mouse testis tissue and analyzing its expression patterns in the undescended testis and various organ tissues as well as in different developmental stages of the mouse testis. The sequencing results of TSEG-2 underwent bioinformatic analyses.
The novel mouse testis gene TSEG-2 was successfully cloned, with full-length sequence of 451 bp. The open reading frame was 267 bp, coding a protein of 88 amino acid residues, and demonstrated to be correct by RT-PCR. The expression of TSEG-2 was high in the mouse testis, regular in the testis cDNA samples of different postnatal days, and down-regulated in the cryptorchidism model. No obvious homology with other mouse cDNA was found for TSEG-2. The GenBank accession number EU079025 was achieved. Function prediction showed that mouse TSEG-2 was probably a soluble non-secretary protein located at chromosome 15qE3, or a nucleoprotein with 2 phosphorylation sites of protein kinase C (PKC) and 1 of casein kinase II (CK2).
A novel mouse testis specific gene TSEG-2 was successfully cloned, which could be down-regulated by cryptorchidism-inducible 17-beta estradiol. This has prepared the ground for further researches on the biological function and expression regulation of TSEG-2.
通过生物信息学方法克隆小鼠睾丸特异性基因TSEG-2。
从小鼠正常睾丸的表达序列标签（EST）在线数据库ZooDDD获取EST。通过dbEST数据库检索其高度同源的EST序列以构建重叠群，并使用生物医学软件Biolign进行拼接。利用GeneScan软件预测基因组序列中的相应外显子和内含子。根据开放阅读框设计引物。应用逆转录聚合酶链反应（RT-PCR）从小鼠睾丸组织克隆该新基因的cDNA，并分析其在隐睾及各种器官组织以及小鼠睾丸不同发育阶段的表达模式。对TSEG-2的测序结果进行生物信息学分析。
成功克隆了新的小鼠睾丸基因TSEG-2，全长序列为451 bp。开放阅读框为267 bp，编码一个含88个氨基酸残基的蛋白质，经RT-PCR验证正确。TSEG-2在小鼠睾丸中表达较高，在不同出生后天数的睾丸cDNA样本中表达规律，在隐睾模型中表达下调。未发现TSEG-2与其他小鼠cDNA有明显同源性。获得GenBank登录号EU079025。功能预测表明，小鼠TSEG-2可能是一种位于15qE3染色体的可溶性非分泌蛋白，或一种具有2个蛋白激酶C（PKC）磷酸化位点和1个酪蛋白激酶II（CK2）磷酸化位点的核蛋白。
成功克隆了一个新的小鼠睾丸特异性基因TSEG-2，其可被隐睾诱导的17-β雌二醇下调。这为进一步研究TSEG-2的生物学功能和表达调控奠定了基础。
