摘要
针对人HER2/neu基因的mRNA序列,设计并构建shRNA干扰载体,经聚合酶链式反应(PCR)及测序鉴定后转染SK-BR-3细胞,并检测HER2/neu mRNA和蛋白抑制情况及SK-BR-3细胞迁移能力的变化.实验结果表明:构建了3个HER2/neu-shRNA真核表达质粒;3个重组质粒均可不同程度下调HER2/neu mRNA和蛋白表达水平,并抑制SK-BR-3细胞的迁移;其中1号重组质粒效应最强,对mRNA和蛋白的抑制率分别为84.65%和74.98%.
To construct the short hairpin RNA （shRNA） interference expression plasmid vectors of human HER2/neu gene,the recombinant plasmid was identified by PCR coupled with sequencing.The vector was transfected into SK-BR-3 cells,HER2/neu mRNA,protein,and the cell migration were assayed.The result indicated that the construction of recombinant plasmid was successful.Transfection of shRNA plasmid significantly down-regulated HER2/neu mRNA and protein; at the same time,the migration ability of SK-BR-3 cell was obviously inhibited.Recombinant plasmid 1 had the strongest effect,with the inhibition ratios of 84.65％ at the mRNA level and 74.98％ at the protein level.
出处
《吉林大学学报（理学版）》
CAS
CSCD
北大核心
2013年第6期1177-1181,共5页
Journal of Jilin University:Science Edition
基金
吉林省自然科学基金(批准号:201015171)
吉林省科技发展计划项目(批准号:201205017)
吉林省医药产业发展专项基金(批准号:YYZX201150-2)
长春市社会发展科技支撑计划项目(批准号:12SF23)
