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Virologica Sinica, 22 (1) : 46-52, 2007
Research Article
Cloning of M Gene and NP Gene of H5N1 Avian Influenza Virus and Immune Efficacy of their DNA Vaccines
Modern Virology Center, State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China
The M and NP gene of H5N1 avian influenza virus (A/chicken/Hubei/489/2004) were amplified by RT-PCR from viral RNA, and cloned into pMD18-T vector respectively. Then the expression plasmid containing M gene (pHM6-m) or NP gene (pHM6-np) was constructed by inserting M or NP gene into the pHM6 eukaryote expression vector, and sequenced. 32 BALB/c mice (6-week-old) were divided into four groups at random. Three groups of BALB/c mice were inoculated with 30 μg of plasmid pHM6-m, 30 μg of plasmid pHM6-np or the mixture of plasmid pHM6-m (15 μg ) and pHM6-np(15 μg) once via the intramuscular route respectively. A group of mice were injected with 100 μl PBS as controls. Two weeks later, all mice were challenged with homologous H5N1 avian influenza virus, and observed in the following 12 days. The survival rates of mice in pHM6-m group, pHM6-np group and mixed plasmids group were 62.5%, 25.0% and 50.0%, respectively. Results showed that effective protection could be provided by either pHM6-m or pHM6-np, but pHM6-m provided a better protective effect than pHM6-np.
  Published online: 20 Feb 2007
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