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

作者信息

Yang Kai-yan, Chen Dao-jin, Li Xiao-rong, Wen Yan-guang

机构信息

Department of General Surgery, Third Xiangya Hospital, Center South University, Changsha, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Jul;26(7):707-8.

PMID:20862802
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.

摘要

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