利用电子差异展示方法克隆人类睾丸高表达新基因SPATA11

Identification of SPATA11,a Novel Gene Highly Expressed in Human Testis by Digital Differential Display

利用NCBI中的电子差异展示(digitaldifferentialdisplay,DDD)软件,比较来自睾丸(包括睾丸癌)与来自其它组织的EST文库,从筛查人类睾丸中高表达而在其他组织中不表达或低表达的差异ESTs入手,成功克隆了一个在人类睾丸中高表达的新基因SPATA11.RT-PCR实验证实其在成人睾丸高表达.序列分析表明该基因含4个外显子,基因组跨越2.6kb,定位于19p13.3.cDNA编码一个含221个氨基酸,相对分子质量为24.5kD的新蛋白.Northern杂交结果显示该基因含有1.1kb大小的唯一转录本,主要在睾丸中强表达,肝脏、肺、卵巢和肾脏中有微弱表达,而其他组织中该基因无表达.

Using the digital differential display program of the National Center for Biotechnology Information, the number of ESTs from human testis and non testis libraries were quantitatively compared and a contig of ESTs which highly expressed in testis libraries was identified.The full length cDNA of this novel gene SPATA11 was deduced and its expression in adult testis was further confirmed by reverse transcriptase polymerase chain reaction (RT PCR).Sequence analysis showed total 4 exons of SPATA11 gene span a 2.6 kb genomic DNA sequence that was mapped to chromosome 19p13.3.The cDNA encodes a novel protein of 221 amino acids with molecular weight of 24.5 kD.Northern blot analyses revealed that SPATA11 mRNA highly expressed in adult testis and faintly expressed in ovary,lung,liver and kidney with one transcript of 1.1 kb.