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Title: cDNA cloning, sequencing, and expression of <img src='/image/spc_char/alpha.gif' border=0>- and <img src='/image/spc_char/beta.gif' border=0>-neurotoxins from Thai-Malayan krait
Authors: Suntrarachun, Sunutcha
Tirawatnapong, Thaweesak
Khunsap, Suchittra
Puempumpanich, Sunanta
Keywords: expression
neurotoxin fusion proteins
recombinant proteins
Issue Date: Jan-2010
Publisher: CSIR
Abstract: <smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="metricconverter"> Amplification products of <img src='/image/spc_char/alpha.gif' border=0>- and <img src='/image/spc_char/beta.gif' border=0>-neurotoxin from Thai-Malayan krait (<i>Bungarus candidus</i>) 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 m<i>M</i> for 24 h. Neurotoxin fusion proteins were used as antigen to generate antibodies in mice. <i>In vitro</i> neutralization indicated that antibody against neurotoxin fusion proteins raised in mice was able to neutralize 2<img src='/image/spc_char/cross.gif' border=0> LD<sub>50</sub> 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 <i>B. candidus</i> venom. </smarttagtype>
Description: 31-37
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.09(1) [January 2010]

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