关键词: AMFR, 表达载体, RNAi, 转染

Abstract: Objective To construct the expression vector of AMFR gene and AMFRsiRNA, and investigate the effect of AMFRsiRNA on the expression of AMFR gene. Methods AMFR gene was amplified by PCR technique and the product of PCR was inserted into the pcDNA3-FLAGC eukaryotic expression vector. Two AMFRsiRNAs were designed by RNAi technique and the AMFRsiRNAs were inserted into the pSliencer 2.1-U6 neo expression vector. Then human embryo kidney 293T cells were co-transfected with the AMFRsiRNA expression vectors and FLAG-tagged AMFR expression vector. The effect of AMFRsiRNAs on the expression of AMFR gene was identified by Western blot. Results The expression vectors of AMFR gene and AMFRsiRNAs were constructed and confirmed by restriction enzyme digestion and DNA sequence analysis. Western blot showed that AMFRsiRNAs could effectively inhibit the expression of AMFR gene. Conclusions The expression vectors of AMFR gene and AMFRsiRNAs were constructed successfully, and the AMFRsiRNAs could effectively inhibit the expression of AMFR gene.

Key words: AMFR, expression vector, RNAi, transfection