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目的克隆一个新的人类睾丸特异表达基因,并分析其组织表达谱。方法运用“数据库消减杂交”(Digital Differential Display,DDD)方法筛选出在人类睾丸组织中显著高表达并代表新基因的克隆重叠群Hs.126780,对该克隆重叠群进行同源性搜索、序列匹配、延伸,获得新基因的cDNA全长序列;生物信息学方法分析和预测新基因结构和功能;多组织RT-PCR实验分析新基因的组织表达谱。结果新基因cDNA全长为1524bp,开放阅读框为1077bp,编码蛋白质由358个氨基酸组成,基因定位于16p13.3,多组织RT-PCR证实新基因为人类睾丸特异性基因。新基因暂命名为:septin 12 transcript variant 2(SEPT12),GenBank登陆号为:EF620906.1。结论利用数据库消减杂交等生物信息学方法和实验相结合克隆新的睾丸特异性基因高效、可行,新基因septin 12 transcript variant 2为人类睾丸特异表达基因。
Objective To clone a new human testis-specific gene and analyze its tissue expression profiles. Methods The cloned contigs Hs.126780, which was highly expressed in human testis tissue and represented a new gene, were screened out by using “Digital Differential Display” (DDD) method. Homology search was performed on the clonal contigs, Sequence matching, extension, obtain the full-length cDNA sequence of the new gene; bioinformatics analysis and prediction of new gene structure and function; multi-tissue RT-PCR analysis of new gene expression profiles. Results The full length cDNA of the new gene was 1524bp. The open reading frame was 1077bp. The encoded protein consisted of 358 amino acids. The gene was located at 16p13.3. Multiplex RT-PCR confirmed that the new gene was a human testis-specific gene. The new gene tentatively named: septin 12 transcript variant 2 (SEPT12), GenBank accession number: EF620906.1. Conclusions The new testis-specific gene was cloned by bioinformatics methods such as database subtractive hybridization. The new testis-specific gene was highly efficient and feasible. The new gene septin 12 transcript variant 2 is a human testis-specific gene.