[Construction and identification of shRNA expressing vectors targeting RhoA gene].

ABSTRACT

AIM: To construct and identify shRNA interference vector targeting RhoA gene. METHODS: Two pairs of single stranded oligonucleotides encoding RhoA siRNA sequence were designed and synthesized. After annealing, its were inserted into vector pGPU6/GFP/Neo, constructed recombinant vectors and then were identified by restrictive digestion and DNA sequencing. LoVo cells were transfected with the recombinant DNA samples by the liposome complex method, and then fluorescence photographs were taken. RESULTS: Enzyme digestion and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pGPU6/GFP/Neo vector. CONCLUSION: The RhoA gene-targeted siRNA and its vector are successfully constructed.