Expression of glutathione S-transferase-cardiotoxin fusion protein in Escherichia coli.
We report here the construction of cardiotoxin V gene, from cobra snake venom (Naja naja atra), by chemically synthesized oligonucleotides and its expression as a glutathione S-transferase-cardiotoxin fusion protein in the inclusion bodies of Escherichia coli. The expression of cardiotoxin fusion protein in protein with a yield of about 35 mg/liter culture was confirmed by highly specific anti-peptide antibodies generated against the unique amino acid residues located at the tip of loop II of cardiotoxin V. Since the fusion protein can be easily treated by CNBr to free the toxin moiety, as revealed by immunoblotting of the cleaved protein, the results provide an avenue for future structural and functional studies of cardiotoxin molecules.
Chi
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LM
Institute of Life Sciences, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Vyas
A A
AA
Rule
G S
GS
Wu
W G
WG
D000595:Amino Acid Sequence; D000818:Animals; D004179:Cobra Cardiotoxin Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D004546:Elapid Venoms / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013329:Structure-Activity Relationship
D004179:Cobra Cardiotoxin Proteins; D004546:Elapid Venoms; D011993:Recombinant Fusion Proteins; C082541:cardiotoxin V; D005982:Glutathione Transferase