Cloning, expression and purification of gussurobin, a thrombin-like enzyme from the snake venom of Gloydius ussuriensis.
Total RNAs were extracted from the venom gland of the snake Gloydius ussuriensis. The cDNA of gussurobin, a thrombin-like enzyme from Gloydius ussuriensis, was cloned and amplified by RT-PCR. Assay of the nucleotide sequence of the cDNA allowed postulation of the complete amino acid sequence for Gloydius ussuriensis, Chinese Viperdae. Its amino acid sequence exhibits significant homology with that of other snake thrombin-like enzymes. The cDNA of gussurobin was inserted into the vector pPIC 9K and expressed successfully in Pichia pastoris, strain GS115. The recombinant gussurobin was separated and purified from 500 ml culture and showed one band on SDS-PAGE.
Yang
Qing
Q
Bioengineering Institute, Dalian University of Technology, Dalian 116012, China.
Hu
Xue-Jun
XJ
Xu
Xiao-Ming
XM
An
Li-Jia
LJ
Yuan
Xiao-Dong
XD
Su
Zhi-Guo
ZG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D018076:DNA, Complementary / Q000032:analysis; D004926:Escherichia coli; D005341:Fibrinolysin / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification; D012911:Snakes
D003435:Crotalid Venoms; D018076:DNA, Complementary; D011994:Recombinant Proteins; D005341:Fibrinolysin; C453734:gussurobin