甲型H1N1流感病毒相变的分子生物学基础研究

Guo Y, Dong J, Wang M

Institute of Virology, Chinese Academy of Preventive Medicine, Bejing 100052, China.

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2000 Mar;14(1):9-13.

OBJECTIVE

To reveal the basis of molecular biology of the phase change of influenza A (H1N1) viruses.

METHODS

Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method, using synthetic oligodeoxynucleotide primers. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03) and Editseq (Version 3.69) software.

RESULTS

No special amino acid on HA1 protein molecule determining the influenza A(H1N1) virus with O or D phase feature was found. However, there are differences of amino acid sequences at -2, -7, 130 and 139 positions between H1N1 viruses isolated in or before end after 1995. It is very interesting that since 1995, two different genetic lineages of influenza A (H1N1) virus HA gene have been cocirculating in men in China. One lineage is no any deletion of amino acid on HA1 protein molecule. The other one has a deletion at 130 position. Their amino acid sequences were compared with those of H1N1 viruses isolated before 1995.

CONCLUSIONS

There is no special amino acid on HA1 domain protein molecule determining the influenza A (H1N1) virus with O or D phase feature was found. Since 1995, two distinguished genetic lineages of influenza A H1N1 virus HA gene have been cocirculating in men in China.

目的

揭示甲型H1N1流感病毒相变的分子生物学基础。

方法

通过逆转录酶将病毒粒子RNA转录为cDNA，用PCR扩增cDNA，纯化PCR产物。然后，使用合成寡脱氧核苷酸引物，通过双脱氧核苷酸链终止法进行RNA序列分析。最后，用MegAlign（版本1.03）和Editseq（版本3.69）软件对测序数据进行系统发育分析。

结果

未发现HA1蛋白分子上有决定O或D相特征的甲型H1N1流感病毒的特殊氨基酸。然而，1995年底及以后分离的H1N1病毒与1995年之前分离的H1N1病毒相比，在-2、-7、130和139位氨基酸序列存在差异。非常有趣的是，自1995年以来，中国人群中甲型H1N1流感病毒HA基因的两个不同遗传谱系同时存在。一个谱系的HA1蛋白分子上没有任何氨基酸缺失。另一个在130位有缺失。将它们的氨基酸序列与1995年之前分离的H1N1病毒的氨基酸序列进行了比较。