ROCK gene shRNA expression vector plasmid and identification of.docVIP

 PAGE \* MERGEFORMAT 11 ROCK gene shRNA expression vector plasmid and identification of Of: Li Xiaojiang deep Yang Yougeng clouds Abstract Objective To construct the expression of Rho kinase (ROCK) specific siRNA recombinant plasmid. Method in accordance with the design principles Elbashir siRNA expression vector and the requirements, the use of Ambion’s online siRNA Tatget finder software, designed with BamH I, Bbs I digested sticky ends, terminate recognition sequence and LOOP loop shRNA, cloned into vector pGPU6/GFP/Neo, ROCK specific siRNA construct the recombinant plasmid, and then restriction enzyme digestion and gene sequencing. ShRNA were designed to successfully cloned into vector pGPU6/GFP/Neo, ROCK specific siRNA construct the recombinant plasmid, restriction enzyme digestion and gene sequencing showed that exactly match the design sequence of the target gene sequence is correct. Conclusion The recombinant plasmid was successfully constructed in vitro or in vivo studies for the future ROCK genes in functional recovery after spinal cord injury provides an effective method. Keywords: vector; RNA interference; ROCK (Rho kinase) RNA interference (RNAi) is a double-stranded RNA mediated mRNA level off in the corresponding post-transcriptional gene expression in the process. With the target gene mRNA transcripts of double-stranded complementary sequences homologous RNA (dsRNA) into cells to generate small interfering RNA (siRNA) combined with RNAi specific enzyme, the formation of RNA-induced silencing complex, with sequence-specific endonuclease activity that specifically degrade the mRNA, resulting in a corresponding lack of functional phenotype. In this study, the vector containing H1 promoter pGPU6/GFP/Neo, for the ras gene superfamily, Rho kinase (ROCK) gene short hairpin RNA (shRNA) targeting ROCK specific shRNA construct the eukaryotic expression vector for the future ROCK in vitro or in vivo gene in functional recovery after sp