Melanin-concentrating hormone receptor 2 gene and identification of siRNA recombinant adenovirus vector.doc

 PAGE \* MERGEFORMAT 13 Melanin-concentrating hormone receptor 2 gene and identification of siRNA recombinant adenovirus vector [Abstract] Objective: To construct Melanin-concentrating hormone receptor 2 (MCHR2) siRNA adenovirus vector and stable expression in Chinese hamster ovary MCHR2’s (CHO) cells, identify their interference effect. Methods: The synthetic siRNA targeting the MCHR2 disturbance sequence, molecular cloning approach and cloned into the shuttle vector pSES  HUS get pSES  HUS  MCHR2siRNA, and with the adenovirus backbone plasmid pAdeasy  1 in BJ5183 bacteria to homologous recombination, to be pAdeasy  SES  HUS  MCHR2siRNA In HEK293 cells, packaged into recombinant adenovirus infection of CHO cells stably expressing MCHR2 lines, RT  PCR and Western Blot Detection of adenovirus on cell MCHR2 expression. Results: MCHR2siRNA adenovirus vector was successfully constructed. MCHR2siRNA adenovirus significantly inhibited the expression of CHO cells MCHR2. Conclusion: Construction of adenovirus Adeasy  MCHR2 siRNA can effectively inhibit the expression of CHO cells MCHR2 for the study of MCHR2 function and its relationship with the relationship between obesity provides an effective tool. [Keywords:] adenovirus containing melanin-concentrating hormone receptor RNA interference 0 Introduction Melanin-concentrating hormone (MCH) is a pivotal role as a neuropeptide can adjust eating, energy metabolism and mood changes. David et al [1] found that MCH receptor MCHR2 may be a candidate gene for obesity in children is one. RNA interference (RNA interference, RNAi) is a double-stranded RNA-mediated, sequence-specific post-transcriptional gene silencing (PTGS) phenomenon, its main biological function is to resist viral infections, transposon transfer to maintain the stability of the genome, removing abnormal RNA, and involved in the regulation of gene expression. our study group has screened against MCHR2 effective interference sequence, we