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| The Prokaryotic Expression and Identification of HN Gene Fragment of Newcastle Disease Virus Strain 7793 |
| LIANG Ying, LIU Jin-ying, FAN Xiao-hui, SONG De-zhi, XIAO Qing, YIN Jun, FENG An-lin, YANG Li, ZHOU Dan-ni, LAI Zhen-pin |
| Guangxi Medical University, Nanning 530022, China |
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Abstract The hemagglutinin-neuraminidase(HN)protein, which is an important spike on Newcastle disease virus (NDV) envelope. It not only has function binding the cell to initiate infection, but also involves in NDV oncolytic effects. HN of NDV serves as a ligand structure for NKp46.NDV-infected tumor cells stimulated NK calls to produce increased amounts of the effector IFN-γ and TNF-α. However, the mechanism about whether HN protein can bind to NKp46 receptor on NK cells and cause TRAIL up-regulation has not yet been clear. The purpose is to obtain HN recombinant protein for further signal transduction researches in NDV oncolytic mechanism. According to the sequence of NDV HN gene available in GenBank, a pair of primers is designed. The target sequence, coding for major antigenic determinants, was amplified by RT-PCR, and was ligated to prokaryotic expression vector pET30a to construct a recombinant expression plasmid pET-HNa. The pET-HNa was transformed to E.coli strain BL21(DE3), and the recombinant HN protein was induced and expressed by IPTG induction. The results of FCM showed that HN not only can bind to NKp46 receptor on resident mice NK cells but also can raise TRAIL production of NK cell. Blocking experiment shows that anti-HN partially inhibit this effect. The results provide basic material for further study on signal transduction in HN stimulated NK cells.
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Received: 19 September 2012
Published: 25 August 2013
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Cite this article:
LIANG Ying, LIU Jin-ying, FAN Xiao-hui, SONG De-zhi, XIAO Qing, YIN Jun, FENG An-lin, YANG Li, ZHOU Dan-ni, LAI Zhen-pin. The Prokaryotic Expression and Identification of HN Gene Fragment of Newcastle Disease Virus Strain 7793. China Biotechnology, 2013, 33(8): 32-37.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2013/V33/I8/32
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