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

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.coli BL21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.coli BL21中表达相对分子质量（Mr）约为59 000的重组融合蛋白，表达量占菌体总蛋白量的30%。

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

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.