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Molecular cloning of a cardiotoxin structural gene from Malayan spitting cobra (Naja naja sputatrix)
M S Yeo1, K Jeyaseelan, M C Chung
1Department of Anatomy, National University of Singapore.
Abstract:
The structural gene and cDNA encoding cardiotoxin in Naja naja sputatrix have been cloned and characterized with a view to study the gene protein relationships and also to produce pure protein in large amounts. Using the polymerase chain reaction on the total RNA isolated from the venom glands, the structural gene (180 bp) has been synthesized and expressed in Escherichia coli to produce a fusion protein with beta-galactosidase. Immunoblotting using polyclonal antibodies raised against the total venom in rabbits demonstrated the presence of cross-reacting proteins in plaques produced by recombinant lambda gt11 phages.