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Title: cDNA cloning, sequencing, and expression of - and -neurotoxins from Thai-Malayan krait
Authors: Suntrarachun, Sunutcha
Tirawatnapong, Thaweesak
Khunsap, Suchittra
Puempumpanich, Sunanta
Keywords: expression;induction;neurotoxin fusion proteins;recombinant proteins
Issue Date: Jan-2010
Publisher: CSIR
Abstract:  Amplification products of - and -neurotoxin from Thai-Malayan krait (Bungarus candidus) were cloned and expressed in TA expression vector. The expressed protein could not be distinguished by SDS-PAGE. Hence, immunoblotting was performed using AntiHis (C-term)-HRP antibody. The antibody could identify the histidine tags at 24 h incubation with 0.02% L-arabinose. To increase the expression level, PCR products were cloned into PCR2.1 cloning vector and pGEX2T expression vector. The optimal condition for protein expression was IPTG induction at 1 mM for 24 h. Neurotoxin fusion proteins were used as antigen to generate antibodies in mice. In vitro neutralization indicated that antibody against neurotoxin fusion proteins raised in mice was able to neutralize 2 LD50 of crude venom. This result provides basic data for the use of the neurotoxin fusion proteins as immunogens in the development of specific antivenoms against the B. candidus venom.
Page(s): 31-37
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.09(1) [January 2010]

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