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Virologica Sinica, 21 (2) : 181-185, 2006
Research Article
Cloning and Expression of Phospholipase A2 Domain of Periplaneta fuliginosa Densovirus in E.coli
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China
The phospholipase A2 functional domain of PfDNV capsid gene VP1 was obtained by
RT-PCR amplification. The amplified fragment was ligated into a pMD18-T vector and sub-cloned into
prokaryotic expression vector pET28a and pET26b. The recombinant plasmid pET28a-PLA and
pET26b-PLA were used to transform E. coli BLl21-codonplus(DE3)-RIL competent cells. After
induction by IPTG, SDS-PAGE indicated that the highly expressed fusion protein was produced. The
fusion protein was purified with Ni-NTA affinity columns and analyzed by Western blot using mouse
anti-His monoclonal antibodies. The results demonstrated that the recombinant PLA2 protein of PfDNV
capsid gene was successfully expressed, which should facilitate further studies on the biological
properties of the enzyme and its function in virus infection.
  Published online: 20 Mar 2006
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