Citation: YU Hai-yang, ZHANG Jia-min, YANG Bo, JIANG Hong, ZHOU Liang, LU Jie, CHENG Wu-guo, LI Dou-lin, HU Yuan-yang*. Cloning and Expression of Phospholipase A2 Domain of Periplaneta fuliginosa Densovirus in E.coli .VIROLOGICA SINICA, 2006, 21(2) : 181-185.

Cloning and Expression of Phospholipase A2 Domain of Periplaneta fuliginosa Densovirus in E.coli

  • Available online: 20 March 2006
  • 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.

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    Cloning and Expression of Phospholipase A2 Domain of Periplaneta fuliginosa Densovirus in E.coli

    • 1. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China

    Abstract: 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.

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