Establishment of transgenic mice with GAP-43 over expression

Objective To establish transgenic mice with over expression of GAP-43 protein,in order to study further on effects and mechanism of GAP-43 in repair of the neural injury.Methods The GAP-43 RNA was extracted from adult C57 mouse and reversely transcripted for obtaining its cDNA.The GAP-43 cDNA was then cloned into the pCEP4-PDGF vector and verified by PCR and sequencing.The plasmid vector pCEP4-PDGF-GAP-43 was transfected into the E.coli DH5α.Transgenic component of pCEP4-PDGF-GAP-43 was therefore constructed and linearized.After purifying,the vector was microinjected into fertilized mouse eggs.These eggs were then transplanted into pseudopregnant mice.The genotype of transgenic mice was identified by PCR.The over expression of GAP-43 protein in the hippocampus of transgenic mice was detected by Western blotting and immunoflourescent staining.Results There were 9 mice showing to have a positive integration of transgenic GAP-43 among 19 experimental mice,by detection of PCR.The Western blotting and immunoflourescence showed that the expression of GAP-43 protein in those transgenic mice was higher than that in the control group.Conclusion The transgenic mouse with GAP-43 over expression was successfully established in the present study.