CAJ | 학술논문

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(expressed sequence tag)筛选大鼠睾丸cDNA文库，获得一个新的cDNA序列-RSD5。RSD5 cDNA全长1556bp,编码176个氨基酸，GenBank接收号为AF146738。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。Northern blot 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。
위료분리、극륭정자발생상관기인，심입료해정자발생적분자궤리，본문이대서고환곡세정관현미분리결합DDRT-PCR적방법，소득적불동구단중차이표체적EST(expressed sequence tag)사선대서고환cDNA문고，획득일개신적cDNA서렬-RSD5。RSD5 cDNA전장1556bp,편마176개안기산，GenBank접수호위AF146738。RSD5기인편마단백서렬함유일개PEST기서，저시단백질쾌속강해적표지。Northern blot 분석결과현시，RSD5재고환조직화뇌조직표체량교고，차재불동발육천령적고환조직중구유현저적표체차이성。RSD5기인적표체특정급기편마단백적PEST기서제시RSD5단백가능재정자발생중구유중요작용。
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.