摘要
将构建保存的重组表达质粒pET-ApxⅠA、pET-ApxⅢA和pPRoAppOML-1用IPTG诱导表达,选择最佳的表达条件为IPTG 1.0 mmol/L,温度为37℃,表达时间为8 h,表达产物用SDS-PAGE鉴定,分别表达41、414、5 ku的蛋白,与预期蛋白大小一致。将表达产物用试剂盒进行纯化,得到较好的目的条带,并对其进行Western blot检测,经过分析,pET-ApxⅠA、pET-ApxⅢA表达产物具有较好的免疫反应性。
The recombinants pET-ApxⅠA,pET-ApxⅢA and pPRoAppOML-1 were constructed by testing laboratory,proteins were proved by SDS-PAGE.Bacterial lyses prepared from 1.0 mmol/L IPTG at 37℃ for 8h induced cultures were loaded directly on to SDS-PAGE.Upon induction,the recombinant pET-ApxⅠA,pET-ApxⅢA and pPRoAppOML-1 proteins with apparent MW of 41,41,45 ku were confirmed.Their MW were the same as expecting proteins.Then The proteins were purified by kit,they were immunoreactive when detected by Western blot analysis.These resulets suggested that purified protein has bioactivity.
出处
《动物医学进展》
CSCD
2007年第12期24-27,共4页
Progress In Veterinary Medicine
基金
"十一五"国家科技支撑计划子课题(BAD06A12)
