摘要
从ConA活化的昆明小鼠脾细胞中提取总RNA,利用RT-PCR方法扩增得到小鼠的IL-10基因。编码序列经测序验证后,将其克隆入表达载体pET-28a,构建IL-10基因的重组原核表达载体,在大肠杆菌(DE3)中诱导表达目的蛋白。SDS-PAGE结果表明,该表达产物的分子量与预期值相符,Western blot结果说明小鼠IL-10基因得到了表达。
The total RNA was extracted from spleen cells of Kunming mouse stimulated by ConA. IL-10 gene of the mouse was amplified by reverse transcription-polymerase chain reaction （RT- PCR）. Coding sequence which was identified by DNA sequencing was cloned into the expression vector PET-28a to construct prokaryotic expression vector. The recombinant protein was expressed in B121 （DE3）. The result of SDS - PAGE showed that the molecular weight of the expressed product was identical to that of prediction. The result of Westernblot demonstrated that the expressed product had reactiongenicity.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2009年第4期434-437,共4页
Journal of Jilin Agricultural University
基金
中国人民解放军医药卫生科技攻关项目(06G138)
