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为了提高人禽流感病毒血凝素HA的表达量,应对流感大流行疫苗的需求,按照人的偏爱密码子将H5N1(A/Anhui/1/2005)流感病毒的HA基因进行优化改造,经全基因合成后插入到真核表达载体pDC315中,构建了真核表达质粒pDC315-Mod·HA;将此质粒和含野生HA基因的真核表达质粒pDC315-Wt·HA分别转染293T细胞,比较HA蛋白的表达量。结果表明:经间接免疫荧光实验及Western blot实验比较和鉴定,密码子优化后,HA蛋白在293T细胞中的表达水平显著提高,为流感大流行疫苗的研究打下了基础。
In order to increase the HA level of the human bird flu virus, the HA gene of H5N1 (A / Anhui / 1/2005) influenza virus should be optimized and reformed according to human preference codons according to the needs of the influenza pandemic vaccine. The gene was inserted into the eukaryotic expression vector pDC315 to construct the eukaryotic expression plasmid pDC315-Mod · HA. The plasmid and the eukaryotic expression plasmid pDC315-Wt · HA containing the wild-type HA gene were transfected into 293T cells respectively to compare HA The amount of protein expression. The results showed that the expression of HA protein was significantly increased in 293T cells after indirect immunofluorescence assay and Western blot, and laid the foundation for the study of influenza pandemic vaccine.