密码子优化的人H5N1流感病毒HA基因在哺乳动物细胞中获得高效表达

Codon Optimization of the H5N1 Influenza Virus HA Gene Gets High Expression in Mammalian Cells

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 improve the expression of human avian influenza virus hemagglutinin （HA） and meet the pandemic influenza vaccine needs, we optimized and synthesized the whole length of HA gene of H5N1 （A/Anhui/1/2005） influenza virus in accordance with the human＇s codon preference, and inserted it to the eukaryotic expression vector pDC315 to construct a eukaryotic expression vector pDC315-Mod. HA. This plasmid and the eukaryotic expression vector pDC315 Wt. HA containing wild HA gene were transfected into 293T cells respectively to compare the expression of HA protein. The results showed that according to the comparison and identification by indirect immunofluorescence assay and Western blot test, the expres- sion of HA protein in 293T cells was significantly improved after codon optimization. This laid a foundation for pandemic influenza vaccine research.