【摘要】 目的:筛选大鼠精子细胞变态相关的cDNA片段。方法:提取两个不同发育阶段的大鼠睾丸总RNA,用mRNA差异显示(mRNA Differential Display)的方法寻找精子细胞变态期特异表达的cDNA片段,并以Northern杂交法进行鉴定。结果:初步找出性成熟大鼠特异表达的cDNA片段。经过测序分析后得到3个同源性低的新cDNA片段,分别命名为DD1-35,DD2-81,DD5-40。以DD5-40为探针,进行Northern杂交,发现此片段只与性成熟大鼠睾丸RNA有杂交信号。结论:表明此片段在精子细胞变态过程中表达,判断其与精子细胞变态相关。更多还原

【Abstract】 Objective To isolate cDNA fragments which express specially in spermiogenesis. Methods for the differ-ence between 7-8-day-old rat testis (contain spermatogonia and apermatocytes)and sexual mature rat lestis (contain spermatogonia , apermatocytes, round spermatids and elongating spermatids ) , we isolated the total RNA from two typies of testes. First, reverse transcription (RT)was carried out to convert the total RNA into cDNA with anchored primer (AP)and arbitrary primer (ARP) ; then, used AP-TMR ( TMR: tetramethylrhodamine) and ARP to get PCR product. After PCR amplification, the fluorescent labelled cDNAs were electrophoresed in 6% denaturing polyacry-lamide gels. Then, the differentially expressed cDNAs were selected and cloned in sequencing vector. New ESTs were found by homology searches and Northern blot was used to analvse one of the new ESTs. Results Five ESTs which named DD1-35, DD2-81, DD3-D, DD4-D DD5-40 had been obtained. Homology searches revealed that DD1-35, DD2-81 , DD5-40 are new cDNA sequence. Northern blot analvsis of DD5-40 confirmed that it was ex-pressed specifically in sexual mature rat testis . Conclusions These results revealed that DD5-40 may be spermiogenesis-related new EST.更多还原