可诱导性表达多种流感病毒HA蛋白稳定细胞系新型构建方法的研究

Rapid construction of stable cell lines which inducible express HA proteins from various influenza virus strains

目的 建立一条创新性的技术路线快速构建可诱导性表达多种甲型流感病毒（IAV）HA蛋白的细胞系.方法 将多种IAV的HA蛋白基因连接到含有PiggyBac转座子位点的Cumate诱导表达系统中,与PiggyBac转座酶质粒共转染HEK293A细胞系,利用嘌呤霉素筛选阳性细胞,并用Cumate诱导相应病毒蛋白表达. 结果 流式细胞仪检测及Western blot检测结果均表明,加入诱导剂Cumate后,获取的细胞系中绝大部分细胞均开始表达相应的病毒蛋白.结论 利用PiggyBac转座子可以高效率的构建诱导性表达IVA HA蛋白的细胞系.

Objective An innovative technique was established to rapidly construct various cell lines that could be induced to express multiple influenza A virus （IAV） proteins.Method The HA protein genes of multiple IAVs were cloned into the Cumate-induced expression system which was positioned between two PiggyBac transposon sites.These HA plasmids were transfected into the HEK293A cell line with the PiggyBac transposase plasmids.The transfected cells were screened with puromycin, and after that the corresponding virus proteins were induced with Cumate.Results The results of flow cytometry and Western blotting showed that the virus proteins were expressed in most of the cells in corresponding lines after the induction of Cumate.Conclusion Cell lines which were inducible to express IVA HA protein can be efficiently constructed by using the PiggyBac transposon system.