大鼠睾丸精子发生中差异表达基因RSD5的克隆与表达分析

Cloning and characterization of a novel rat gene RSD5 differentially expressed in spermatogenesis

为了分离、克隆精子发生相关基因 ,深入了解精子发生的分子机理 ,本文以大鼠睾丸曲细精管显微分离结合DDRT PCR的方法 ,所得的不同区段中差异表达的EST(expressedsequencetag)筛选大鼠睾丸cDNA文库 ,获得一个新的cDNA序列 RSD5。RSD5cDNA全长 15 5 6bp ,编码 176个氨基酸 ,GenBank接收号为AF146 738。RSD5基因编码蛋白序列含有一个PEST基序 ,这是蛋白质快速降解的标志。Northernblot分析结果显示 ,RSD5在睾丸组织和脑组织表达量较高 ,且在不同发育天龄的睾丸组织中具有显著的表达差异性。RSD5基因的表达特征及其编码蛋白的PEST基序提示RSD5蛋白可能在精子发生中具有重要作用。

For the understanding of the molecular mechanism of spermatogenesis, the method of DDRT PCR(differential display reverse transcription polymerase chain reaction) combined with segmentation technique of the seminiferous tubule was adopted to isolate and identify the differentially expressed genes in spermatogenesis Three ESTs were used as the probes to screening the rat testis cDNA library A cDNA named RSD5 was isolated from rat testis cDNA library It is 1556bp in length, coding a protein of 176 amino acids with the GenBank accession number AF146738 The encoding protein of RSD5 cDNA has a potential PEST motif, which being a marker related to the rapid degradation Northern blot analysis showed that the RSD5 gene was widely expressed in all of the eight examined tissues, whereas testis and brain presented a higher expression The transcription of RSD5 mRNA in testis was detectable in 6,18,21,40 days after birth and reaches the highest level in 60 days testis of adult rat The function of RSD5 in spermatogenesis remains further investigation