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


CONCLUSIONS
The RhoA gene-targeted siRNA and its vector are successfully constructed.
Yang
Kai-yan
KY
Department of General Surgery, Third Xiangya Hospital, Center South University, Changsha, China. soskinghero@126.com
Chen
Dao-jin
DJ
Li
Xiao-rong
XR
Wen
Yan-guang
YG
D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D018390:Gene Targeting / Q000295:instrumentation* / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D020742:rhoA GTP-Binding Protein / Q000235:genetics* / Q000378:metabolism
D034741:RNA, Small Interfering; D020742:rhoA GTP-Binding Protein