IPTG诱导浓度、时间对AsiaⅠ型口蹄疫病毒VP1蛋白表达的影响

Effect of Induced Concentration and Time of IPTG on the Expression of VP1 Protein of FMDV Type Asia Ⅰ

将口蹄疫病毒VP1基因插入原核表达载体pET-28a中,转化至大肠杆菌BL21。用不同的IPTG浓度与不同诱导时间诱导菌株表达VP1蛋白,分析不同IPTG浓度及不同诱导时间对蛋白表达量的影响,以确定最佳表达条件。结果表明,在一定范围内,随IPTG浓度的增高,表达量并不会随之增大;而在菌体对数生长期内,随诱导时间的延长,表达量会随之增大。最后确定当IPTG浓度为3 mmol/L,诱导表达到最长时间9 h时,VP1蛋白表达量最大。

The VP1 gene of FMDV type Asia I was inserted into the prokaryotic expression vector pET-28a, and then transformed into BL21 cells. The recombinant plasimd was induced with different concentration and time of IPTG to express the VP1 protein. The better expression condition was confirmed by analyzing the effect of different concentration and time of IPTG on the expression product. The experimental result indicated that the expression product had not been increased with adding of the concentration of IPTG in definite scope, but the expression output had been increased with the extension of time during the period of logarithmic growth of bacteria. The maximal quantity of expression of VP1 protein was confirmed when the concentration of IPTG was 3 mmol/L and the induced time was 9 hours.