阴道毛滴虫沉默信息调节因子2同源基因的原核表达载体构建及表达  

Construction of Prokaryotic Expression Vector for Trichomonas vaginalis Silent Information Regulator 2 and Its Expression

作　　者：张可浩[1] 黄丽霞[1] 傅玉才[2] 章家新 

机构地区：[1]温州医学院附属台州医院,台州317000 [2]汕头大学医学院,汕头515041 [3]厦门市仙岳医院,厦门361012

出　　处：《中国寄生虫学与寄生虫病杂志》2006年第3期244-246,共3页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：提取阴道毛滴虫(T.vaginalis)LX006细胞株传代培养细胞总RNA,RT-PCR扩增Tv-Sir2-like全长cDNA链,扩增产物连接到T-A克隆载体并转化大肠埃希菌(E.coli)JM109。提取重组克隆质粒,并用限制性内切酶EcoRⅠ和PstⅠ进行双酶切,将目的基因定向克隆到原核表达载体pET-41b,并在E.coliBL21中经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达。结果显示目的基因定向克隆于原核表达质粒pET-41b获得了表达重组子,IPTG诱导该原核表达载体在E.coliBL21中表达相对分子质量(Mr)约为59000的重组融合蛋白,表达量占菌体总蛋白量的30%。Total RNA was isolated from Trichomonas vaginalis and Tv-Sir2-like cDNA was amplified by RT-PCR and cloned into pGEM-T Easy plasmid. A fragment of Tv-Sir2-like eDNA was subcloned into the expression vector pET-41b and expressed in E.coli BL21 with induction of IPTG. The full-length of Tv-Sir2-like eDNA was cloned and sequenced. The prokaryotic expression system of pET-41b/Tv-Sir2-like was constructed. The fusion protein of Tv-Sir2-like was expressed in E.coli BL21, occupying 30% of the total bacterial protein after being induced by IPTG for 5 h. SDS-PAGE analysis showed that the fusion protein was about Mr 59000. The recombinant protein of Tv-Sir2-like is efficiently expressed in E.coli BL21.

关 键 词：阴道毛滴虫 沉默信息调控因子2 同源基因 基因克隆 原核表达 

分 类 号：R382.211[医药卫生—医学寄生虫学]