Cloning and functional expression of a synthetic gene encoding huwentoxin-I, a neurotoxin from the Chinese bird spider (Selenocosmia huwena).
Cloning and functional expression of a synthetic gene encoding huwentoxin-I, a neurotoxin from the Chinese bird spider Selenocosmia huwena. A gene encoding huwentoxin-I, a peptide neurotoxin consisted of 33 amino acid residues from the venom of the Chinese bird spider Selenocosmia huwena, was designed, synthesized and expressed in Escherichia coli as a hybrid protein fused with glutathione S-transferase at the N-terminal. The fusion protein was purified by GSH-Sepharose 4B affinity column chromatography and cleaved by thrombin to release the toxin peptide. The amino acid sequence of the recombinant toxin was consistent with the designed one by sequence determination and MALDI-TOF mass analysis, suggesting that the recombinant huwentoxin-I produced the same expression product as the native one. After reduction and renaturation, the biological activity of the recombinant toxin was identical with that of the native huwentoxin-I by electrophysiological method.
Li
M
M
Department of Biology, Hunan Normal University, Changsha, People's Republic of China.
Li
L Y
LY
Wu
X
X
Liang
S P
SP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009469:Neuromuscular Junction / Q000187:drug effects; D009498:Neurotoxins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D013111:Spider Venoms / Q000235:genetics*; D009435:Synaptic Transmission / Q000187:drug effects
D009498:Neurotoxins; D011994:Recombinant Proteins; D013111:Spider Venoms