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Acta Prataculturae Sinica ›› 2014, Vol. 23 ›› Issue (4): 138-145.DOI: 10.11686/cyxb20140417

• Orginal Article • Previous Articles     Next Articles

Vector construction and over-expression of an avian influenza virus hemagglutinin (HA) in transgenic Lotus corniculatus

ZHANG Wei-wei1,2,YUAN Bei2,ZHANG Zhan-lu3,ZHAO Yang-min3,XU Bing-liang1,WU Yan-min2   

  1. 1.College of Grassland Science,Gansu Agricultural University,Lanzhou 730070,China;
    2.Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China;
    3.Shenzhen Nongke Group Corporation,Shenzhen 518040,China
  • Received:2014-01-15 Online:2014-08-20 Published:2014-08-20

Abstract: A safe and efficient plant expression vector,HA-LTB,which added double 35S promoter,Ω enhancer,endoplasmic reticulum retention signal KDEL sequence and extended poly (A) sequence was constructed. Twenty one transgenic Lotus corniculatus plants were obtained by the Agrobacterium-mediated method and the results of PCR and RT-PCR showed that the HA-LTB gene had been obtained from the plant genome. ELISA results of the B subunit of Escherichia coli heat-labile enterotoxin (LTB) showed that LTB could be expressed in transgenic L. corniculatus and had immunogenicity. It was concluded that hemagglutinin (HA) which was fused with LTB had immune activity. The highest expressions of LTB and HA were 0.086 μg/mg (crude protein extracts) and 0.25 μg/mg (crude protein extracts) respectively which has approached advanced international standards. A high level exogenous antigen protein expression platform which was a basis for research on the H5N1 avian influenza vaccine was established by optimizing the expression vector.

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