title: [Construction of prokaryotic expression vector for Trichomonas vaginalis silent information regulator 2 and its expression].
abstract: 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 cDNA 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 cDNA 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.
authors.affiliation: Taizhou Hospital of Zhejiang Province, Taizhou 317000, China.
authors.forename: Ke-hao
authors.initials: KH
authors.lastname: Zhang
authors.forename: Li-xia
authors.initials: LX
authors.lastname: Huang
authors.forename: Yu-cai
authors.initials: YC
authors.lastname: Fu
authors.forename: Jia-xin
authors.initials: JX
authors.lastname: Zhang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D037761:Sirtuins / Q000235:genetics*; D014246:Trichomonas vaginalis / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D037761:Sirtuins