Establishment of BIGH3~(R555W)-mutation transgenic mice

Objective To establish an animal model of BIGH3~(R555W)mutation.Methods The fulllength human BIGH3 with R555w mutation was inserted into the IRES-EGFP vector under the control of phosphoglycerate kinase(Pgk) promoter.The recombinant transgenic fragment was constructed,linearized and purified,men microinjected into fertilized mouse eggs.These eggs were transplanted into pseudopregant mice.The genotypes of transgenic founders were identified by PCR.The expression of mutant BIGH3~(R555W)was confirmed by RT-PCR and Western blotting.Results Transgenic C57 mice were obtained by microinjection.PCR results showed 4 out of 35 mice were integrated.The mutant BIGHS~(R555W) was overexpressed in the cornea of transgenic mice compared to wild-type mice as demonstrated by RT-PCR and Western blotting analysis.Conclusion BIGH3~(R555W) mutation transgenic mouse model has been successfully established by pronuclear microinjection,the transgenic mice would provide a animal model for study of pathogenesis of corneal dystrophy.