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

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.