阴道毛滴虫病毒RDRP基因的克隆与分析

Cloning and sequence analysis of the RNA-dependent RNA polymerase(RDRP) gene in Trichomonas vaginalis virus

目的对阴道毛滴虫病毒RNA依赖的RNA聚合酶(RDRP)基因的克隆,以便探讨其功能。方法提取阴道毛滴虫总核酸为模板,根据所克隆的阴道毛滴虫病毒部分序列和GenBank中发表的阴道毛滴虫病毒TVV-T1序列设计一对引物,经RT-PCR得到与预计大小一致的PCR特异性产物,将其克隆到pMD-18T载体后测序,并与GenBank中核苷酸序列进行同源性搜索与分析。结果目的基因片段长度为2271bp,与阴道毛滴虫病毒T1株(U08999)的同源性最高为85.2%。结论国内首次成功克隆出阴道毛滴虫病毒RNA依赖的RNA聚合酶(RDRP)基因序列。

To clone and analyze the RNA-dependent RNA polymerase（RDRP）gene in Trichomonas vaginalis virus in order to investigate its function, a pair of primers was designed according to the cloned partial sequence and the published sequences of T vaginalis virus in GenBank and the total nucleic acid of T vaginalis was extracted and used as template for the amplification of the viral capsid genome by using RT-PCR assay. The specific PCR product was cloned to vector pMD-18T,sequenced and then compared with the nucleotide sequences available in GenBank to analyze their homologies. It was demonstrated that the length of the target gene fragment was 2271 bps and the homology with that of the T1 stain（U08999）in GenBank was rather high（8 2.5 %）. It seems that the nucleotide sequence of the RNA-dependent RNA polymerase gene in T vaginalis virus was first successfully isolated.