Cloning and Expression
of α-Bungarotoxin Gene
DUAN Hai-Qing, ZHANG Zhao-Shan*, LI Shu-Qin and
HUANG Cui-Fen
( Beijing Institute of Biotechnology, Beijing 100071, China )
Abstract The genetic engineering
methods to produce the snake neurotoxin obviously have potential advantages
over the traditional methods of extracting neurotoxin from snakes. This work
was to study the expression and the feasibility of scaled production of snake
neurotoxin in the routine expression systems such as E.coli with the α-bungarotoxin
as an example. First, on the basis of the reported amino acid sequence of α-bungarotoxin,
DNA sequence of α-bungarotoxin was deduced and four partially complementary
oligonucleotide fragments were designed. The coding region of α-bungarotoxin
was obtained by renaturing the DNA fragments, nick filling-in, ligation and
PCR. The coding region of α-bungarotoxin was cloned into plasmids pGEX-2T,
named pDZ04, and transformed E.coli BL21 in order to study the
expression of α-bungarotoxin gene. The results of SDS-PAGE analysis showed that
the recombinant plasmid pDZ04 could express efficiently in BL21, and the fusion
protein took up about 30%―40% of total bacterial protein. Finally, The
biological activity of the recombinant α-bungarotoxin and the fusion protein
was studied with the natural α-bungarotoxin purified from the snake venom as
control, ELISA results showed that they had similar antigenicity.
Key Words Snake neurotoxin； α-bungarotoxin； gene expression
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