Abstract
The cDNA encoding α-bungarotoxin (α-BuTx) was constructed from the cellular RNA isolated from the venom glands of Bungarus multicinctus by RT-PCR. The primers deduced from the N-, C-terminal amino acid residues and the conserved middle segment of the sequence, i.e., R25KMWC29 in α-BuTx could specifically amplify cDNAs encoding amino acid residues from 1 to 29 and 25 to 74 of α-BuTx. The complete cDNA was obtained by linking sufficient amounts of the two cDNA fragments by PCR. The amino acid sequence translated from the α-BuTx cDNA sequence was identical to that of native toxin. The PCR product was then cloned into pGEX vector and transformed in DH5 E. coli strain. The fused protein expressed in host strain exhibited the same antigenicity as native α-BuTx. This communication is the first report for the cloning and expression of a long α-neurotoxin from the venom glands.
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