CAJ | 학술논문

为了拯救出H3N2亚型流感病毒的重组新病毒,利用反向遗传学手段,8个质粒共同转染293T细胞,包装带有双点突变（第119位氨基酸由E突变为V,第222位氨基酸由I突变成L）、单点突变和野生型的流感病毒H3N2;在MDCK细胞中传代包装成的H3N2病毒。结果表明,野生流感病毒H3N2和其突变株包装成功,第119位氨基酸单点突变型滴度与野生型的相近,而第222位氨基酸单点突变和双突变型滴度要比野生型的低。本实验成功证明了H3N2神经氨酸酶上两个位点（第119和222位氨基酸）对病毒包装及复制起到关键的作用,且NA上第222位氨基酸相对于第119位氨基酸显得要更重要。
위료증구출H3N2아형류감병독적중조신병독,이용반향유전학수단,8개질립공동전염293T세포,포장대유쌍점돌변（제119위안기산유E돌변위V,제222위안기산유I돌변성L）、단점돌변화야생형적류감병독H3N2;재MDCK세포중전대포장성적H3N2병독。결과표명,야생류감병독H3N2화기돌변주포장성공,제119위안기산단점돌변형적도여야생형적상근,이제222위안기산단점돌변화쌍돌변형적도요비야생형적저。본실험성공증명료H3N2신경안산매상량개위점（제119화222위안기산）대병독포장급복제기도관건적작용,차NA상제222위안기산상대우제119위안기산현득요경중요。
In order to save the reorganization of a new virus of the H3N2 influenza virus,H3N2 wild type and three mutation type(E119D+I222L,E119D and I222L) viruses were generated by reverse genetics;The reverse genetics viruses were stable after five passages in MDCK cells.Successfully generated H3N2 wild type and mutation type viruses;The wild type had a similar virus titer with the mutation type E119D;The virus titer of E119D+I222L and I222L type were lower than the wild type.These results indicated that double mutation(E119D+I222L) type H3N2 could induce a high level of oseltamivir resistance.NA and the first 222 amino acids compared to 119 amino acids seem to be more important.