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目的 阐明甲 1(H1N1)亚型毒株相变异的分子生物学基础。方法 病毒RNA经逆转录合成cDNA ,利用聚合酶链反应 (PCR)进行扩增 ,产物纯化 ,采用双脱氧链末端终止法进行核苷酸序列测定 ,最后用DNASTAR公司出品的序列分析软件MegAlign(1 0 3版 )和Editseq(3 6 9版 )对核苷酸序列进行分析。结果 见不到“O”、“D”相毒株HA1蛋白分子间有特殊氨基酸的差异。但 1995年前后毒株在 - 2 ,- 7,130和 139位存在有差别。很有意义地发现了 ,自 1995年以来 ,在我国人群中同时流行着两种HA基因不同的H1N1亚型流感病毒 :一种与 1995年之前所分离到的H1N1病毒相比较 ,其HA1区蛋白分子上氨基酸没发生任何丢失 ,另一种在第 130位丢失一个氨基酸。结论 未发现H1N1亚型病毒“O”、“D”相毒株HA1区蛋白分子间有特殊氨基酸的差异。发现了自 1995年以来 ,我国人群中同时流行着两种HA基因不同的H1N1亚型毒株。
Objective To elucidate the molecular biology basis of the phase change of subtype A (H1N1) strains. Methods The viral RNA was reverse transcribed into cDNA and amplified by polymerase chain reaction (PCR). The product was purified and the nucleotide sequence was determined by dideoxy chain termination method. Finally, the DNA sequencing software MegAlign (1 0 3 Edition) and Editseq (399 Edition) for nucleotide sequence analysis. As a result, there was no difference in the special amino acids between the “O” and “D” phase HA1 protein molecules. However, there were differences between 1995 and the strains at - 2, - 7, 130 and 139. It has been found to be of great significance that since 1995, two H1N1 subtype influenza viruses with different HA genes are prevalent in our population: one is a subtype of HA1 protein, compared with the H1N1 virus isolated before 1995 There was no loss of amino acids on the molecule, and the other lost an amino acid at position 130. Conclusion No specific amino acid difference was found between the HA1 region of the H1N1 subtype virus and the “D” phase strain. Found that since 1995, two subtypes of H1N1 isolates with different HA genes are prevailing in our country.