China Animal Husbandry and Veterinary Medicine ›› 2019, Vol. 46 ›› Issue (10): 2860-2866.doi: 10.16431/j.cnki.1671-7236.2019.10.006

• Biotechnology • Previous Articles     Next Articles

Fusion PCR of Chicken Infectious Bursal Disease Virus VP2 and Helicobacter pylori Ferritin Fusion Gene and Construction of Its Eukaryotic Expression Vector

ZHU Yanping1, HE Yong1, LIU Jia1,2, XING Ruilin1, CHANG Lekai1, LI Runzhi1,3, YUE Feng1, WU Yuping1, LI Peng1, SUN Guopeng1, ZHANG Yanfang1, WAGN Xuannian1,2,3   

  1. 1. Center for Biotechnology Research, College of Life Science and Technology, Xinxiang University, Xinxiang 453003, China;
    2. School of Life Science, Zhengzhou University, Zhengzhou 450000, China;
    3. College of Animal Sciences, Henan Institute of Science and Technology, Xinxiang 453003, China
  • Received:2019-04-29 Online:2019-10-20 Published:2019-10-21

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Abstract: In order to obtain the recombinant protein of VP2 protein of infectious bursal disease virus (IBDV) with high activity,the fusion of IBDV VP2 protein and self-assembled Helicobacter pylori (Hp) ferritin gene were amplified by fusion PCR.Two pairs of primers were designed and synthesized according to the DNA sequence of mature VP2 protein gene and the base sequence of Hp ferritin.The fusion gene fragment VP2-Fe was amplified by fusion PCR.The target gene was cloned into pMD18-T vector and the positive recombinant plasmid was screened.Then the target gene was subcloned into eukaryotic expression vector pPICZaC and transformed into competent cells of Escherichia coli DH5α,the positive expression plasmids were screened.The results showed that the fusion gene VP2-Fe with the length of 1 824 bp was amplified by two rounds of PCR and subcloned to pMD18-T vector.Sequencing results showed that there was no mutation in the fusion gene.The positive plasmid was named as pMD-VP2-Fe.The target gene (1 824 bp) was recovered by double digestion and subcloned to pPICZaC.The expected size fragment was identified by PCR and double digestion.The positive expression plasmid was named as pPICZaC-VP2-Fe.This study laid a foundation for the later use of eukaryotic expression vector to obtain its fusion recombinant protein.

Key words: IBDV; VP2 gene; Helicobacter pylori (Hp); ferritin; fusion PCR

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