[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 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.
Zhang
Ke-hao
KH
Taizhou Hospital of Zhejiang Province, Taizhou 317000, China.
Huang
Li-xia
LX
Fu
Yu-cai
YC
Zhang
Jia-xin
JX
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*
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D037761:Sirtuins