Research For Expression Of Somatostatin Receptor (SSTR5) Gene In Mice By ShRNA Interfering

Animal growth controll by growth hormone(GH),including growth hormone releasing factor(GRF)and somatostatin(SS).Somatostatin(SS),a growth hormone secreting pituitary inhibition of neuropeptides,widely distributed in the central and peripheral nervous system of vertebrates,pancreas and gastrointestinal tract,such as the structure of a class-related peptide.SS is the biological function of a variety of G-protein coupled with the five kinds of somatostatin receptor subtypes in different SSTR achieved.Five kinds of different subtypes SSTR mainly distributed in the brain,pancreas,pituitary and other organizations in their respective functions also have some differences.Endogenous or exogenous to the double-stranded RNA into cells,and the homologous mRNA by the RNA degradation,so that the corresponding genes have been restrained.This is caused by RNA interference gene silencing.The method has become so specific gene inactivation as an effective method . SS inhibit gene expression in vivo SS reduce the secretion of GH can be a corresponding increase in the concentration of the final performance for the lower metabolism,thus help promote animal growth purposes.SS would be to reduce the concentration of animals in complex feedback mediation, and SSTRs may play a key role.Against SSTRs undertake a study on the role in the reduction of the SS,is likely to be a better result.Pituitary cells Secretions SS-28 expression level higher than the that of SS-14 and SS-28 has the highest affinity with SSTR5,the present study was to explore application of RNA interference technology to SSTR5-induced gene silencing, thereby using RNA interference inhibit gene expression provide the new SS method.This study uses successfully constructed the SSTR5 (NM₀11425) targeting siRNA expression vector stability of the DNA in cells transfected with SSTR5 inhibited gene expression.Through the transfected cells by RT-PCR,fluorescence quantitative PCR,fluorescence microscopy and ELISA transfected cell lysis in SSTR5 concentration of three SSTR5 siRNA to contain the size of the role of gene expression. Fluorescence quantitative PCR analysis of variance showed that different samples ofβ-actin group there was no significant difference (P>0.05),can be considered different samplesβ-actin expression of the same;transfection pSSTR5-1,pSSTR5-2,pSSTR5-3 in the group relative to the control group SSTR5 initial DNA with significant differences (P<0.05).pSSTR5-1, pSSTR5-2, pSSTR5-3 inhibition rates were 64.20%, 67.36%, 75.64%. ELISA test results,the SPSS12.0 software to do one-way analysis of variance, transfection pSSTR5-1,pSSTR5-2,the group pSSTR5-3 protein expression between SSTR5 no significant difference (P>0.05), but compared to the blank control,pSSTR5-1,pSSTR5-2,the group pSSTR5-3 protein expression of SSTR5 significant difference (P<0.05), or transfected pSSTR5-1, pSSTR5-2,pSSTR5-3 group SSTR5 expression level was significantly lower than empty cells control group (P<0.05), the results shows that the three siRNA inhibiting SSTR5 expression have different levels and inhibition rate were 54.42%, 59.12%, 63.43% The research design three siRNAs targeting of gene SSTR5 in mice,through transfection of pituitary cells.Then detect levels of mRNA and protein expression SSTR5.Results showed that the three SSTR5 siRNA can effectively inhibit gene expression.This is a way that using of RNAi technology to raise animals lay the foundation for growth performance.