Expression of gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis, in Escherichia coli.
Gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis, was expressed in Escherichia coli using expression vector pET-32a(+). The gene was expressed under T7 promotor with a fusion partner of Thx.Tag and a 6xHis.Tag at its 5' terminal. After induction by IPTG for 6 h, the recombinant enzyme was expressed in the cytoplasm. Expression at 25 degrees C gave twice the amount of recombinant gloshedobin in cytoplasm than at 37 degrees C.
Yang
Qing
Q
Bioengineering Institute, Dalian University of Technology, Dalian 116012, PR China.
Li
Min
M
Xu
Jianqiang
J
Bao
Yongming
Y
Lei
Xuyu
X
An
Lijia
L
D000818:Animals; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005340:Fibrinogen / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature; D013917:Thrombin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014757:Viper Venoms / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D017819:Viperidae / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D014757:Viper Venoms; D005340:Fibrinogen; D013917:Thrombin