The Preparation, Detection And Preliminary Analysis Of Phenotype Of The MiRNA-378Transgenic Mice

miRNA play a very important role in regulating the development of muscle in differentstages,meanwhile shows great effects on cell proliferation and differentiation. miRNA-378over-expression vector was constructed in this study and transfected porcine embryo fibroblast（PEF）to verify the expression efficiency of the vector in a cell level.Integration efficiency and expressionefficiency of transgenic mice produced by pronuclear injection technique were analyzed by molecularbiological methods, and the biological function of miRNA-378was analyzed preliminarily in transgenicmice.miRNA-378precursor sequence fragments were amplified from sus scrofa genomic DNA andrecovered by XhoI digestion to connect to pEGP-miR vector. Recombinant vector pEGP-miR-378wassuccessfully constructed to transfect porcine embryo fibroblast（PEF）.The transfection efficiency andthe efficiency of gene expression by fluorescence observation; the expression activity of miRNA-378inthe cells was determined by real-time quantitative PCR（QRT-PCR）.It turned out that pEGP-miR-378recombinant vector was successfully constructed; The transfection cells showed higher transfectionefficiency and higher gene expression efficiency; QRT-PCR results showed that the expressions ofmiRNA-378in experimental group were significantly increased significant（p<0.01）compared to thecontrol group; The results provide technology platform for the preparation of transgenic animals to findthe functions of miRNA-378.miRNA-378over-expression transgenic mice were produced by pronuclear injection followingcombing with the PCR and Southern blot to detect positive transgenic mice for Founder mice. Theoffspring of mice propagation was based on founder mice using full-sib mating,48transgenic mice in77was detected by PCR and Q-PCR and transgenic positive rate was62.3%, The inheritance ofexogenous genes followed the laws in the chromosomes; The transgenic mice were detect the greenfluorescence under blue light detected by vivo imaging technology; Body weight of mice was recordedweekly from weaning（3weeks）to the first10weeks, it showed that the growth rate and weightincreased in some pedigrees of transgenic mice compared with non-transgenic mice pedigrees. Theestablishment of the model provides a way to obtain transgenic mice with relatively faster growth rateand gaining weight, and it has laid a good foundation for the study of miRNA-378biological functionsin future.