阴道毛滴虫cDNA文库的构建和鉴定

Construction and Identification of cDNA Library of Trichomonas vaginalis

目的 :构建阴道毛滴虫(Trichomonasvaginalis,Tv)全长cDNA表达文库 ,筛选全长目的基因 ,研究与其细胞周期相关基因的结构和功能。方法 :从Tv中提取总RNA ,用ClonTech公司的SMARTTMcDNA文库试剂盒构建其全长cDNA表达文库。结果 :所建文库的初始滴度为6 78× 10 6 pfu mL ,经一次扩增后的滴度为 1 6 8× 10 9pfu mL ,重组率 >98%。结论 :成功构建Tv全长cDNA文库 。

Objective:To construct a full-length cDNA library of Trichomonas vaginalis(Tv)for screening genes involved in the cell cycle.Method: Using“SMART cDNA Construction Kit',cDNA was synthesized with the total RNA template isolated from Tv in vitro and the cDNA library was constructed in the phage λ TripIEx2 vector.Results:The primary titer of the constructed library was 6.78×10 6 pfu/mL,amplified library of 1.68×10 9 pfu/mL.The recombinant efficiency was more than 98%.Conclusion: The full-length cDNA library of Tv is constructed successfully.The genes of our interest have been screened. [