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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (09): 19-23    DOI: Q78
    
Genetic Stability and Titer Determination of Recombinant Adenovirus Expressing HA Gene of Strain H5N1 of Avian Influenza Virus
WANG Qing1,HU Jian-he1,XU Yan-zhao2
1.Henan Institute of Science and Technology, Xinxiang 453003,China
2.Chinese Academy of Agricultural Sciences, Shanghai Veterinary Institute, Shanghai 200241,China
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Abstract  

The focus is to evaluate the genetic stability and titer determination of the recombinant adenovirus (pAd-H5) expressing HA gene of H5N1 Avian Influenza virus (AIV). The recombinant adenovirus pAd-H5 had been serially passaged for 20 passages in 293 cell line. The passages of 5,10,15 and 20 were chosen to be identified by PCR and sequence; Using GFP rapid monitoring method to test the viral titer of pAd-H5, passage 20. Furthermore, the analysis supported that HA gene was amplified correctly, without any changes on the nucleotide in passages of 5, 10 and 15. While there was only one point mutation on HA gene (417 A→G) in passage 20, but this point mutation did not lead to amino acid changes (samesense mutation). The titer of pAdH5 in 293 cell line identified by GFP rapid monitoring method was 108.75pfu/0.1ml. All the results showed that the pAd-H5 has good genetic stability within 20 passages, and the titer was relativly high.

Key words： Avian Influenza virus (AIV)      Recombinant adenovirus (pAd)      Genetic stability      Titer determination     
Received: 22 February 2010      Published: 25 August 2010
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WANG Jing
HU Jian-He
XU Pan-Shao
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WANG Jing, HU Jian-He, XU Pan-Shao. Genetic Stability and Titer Determination of Recombinant Adenovirus Expressing HA Gene of Strain H5N1 of Avian Influenza Virus. China Biotechnology, 2010, 30(09): 19-23.

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https://manu60.magtech.com.cn/biotech/Q78     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I09/19

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