Molecular Cloning and
Expression of the cDNA
for Disintegrin from Agkistrodon acutus
FAN Chun-Yang, QIAN You-Cun， SHEN Yan, YANG Sheng-Li and
GONG Yi
( Shanghai Research Center of Biotechnology, the Chinese Academy of
Sciences, Shanghai 200233, China )
Abstract In order to clone novel and
valuable genes from natural resources and develop genetic engineering drugs,
total RNA was extracted from the venom gland of Agkistrodon acutus by
one step method. The cDNAs for low molecular weight metalloproteinase were
amplified by RT-PCR and cloned into the pGEMT vector. Their sequence were
determined. One of the deduced amino acid sequences included a metalloproteinase
and a disintegrin (acugrin) at the carboxyl terminus. This result further
confirms that metalloproteinase and disintegrin are derived from the proenzyme
of snake venom metalloproeinase. Acugrin exhibits a certain amino acid homology
with other disintegrins from snake venoms. Acugrin has four disulfide bonds and
a consensus motif： RGD. The cDNA for the acugrin was cloned
into the plasmid pT7ZZa and expressed in E.coli. ZZ-acugrin fusion
protein could strongly inhibit aggregation of platelet induced by ADP and the
IC50 was about 4 nmol/L.
Key Words disintegrin； Agkistrodon acutus； fusion expression； anti-thrombus
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