Expression of NA Gene of Avian Influenza Virus Subtype H5N1 in Escherichia coli and Purification of the Expressed Product

doi:10.7668/hbnxb.2008.05.023

Abstract

Abstract: The neuraminidase(NA)gene of Avian intluenza virus(AIV)subtype H5 was amplified by PCR according to the sequence published on GenBank.It was cloned into the expression vector-PET-32a.The protein encoded by this gene was expressed in Escherichia coli strain BL21(DE3)and was characterized by SDS-PAGE and western blotting analysis as a 61 kDa protein with immunogenicity.The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation.The results indicated that this protein could be specifically recognized by H5 subtype AIV antiserum.The assay has good specificity to H5 AIV.The ELISA result demonstrated that the purified protein could be used to detect the antibody against NA of AIV subtype H5N1 and had good sensitivity.

Key words: Avian influenza virus, Subtype H5, Neuraminidase (NA), Prokaryotic expression

CLC Number:

S858.31

ZHANG Gai-ping, LI Qing-zhou, HAO Hui-fang, ZHOU Ling, WANG Li, YANG Su-zhen, LUO Jun, FENG Li-li. Expression of NA Gene of Avian Influenza Virus Subtype H5N1 in Escherichia coli and Purification of the Expressed Product[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(5): 107-110. doi: 10.7668/hbnxb.2008.05.023.

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