[Fine mapping of mutant gene related corneal opacity mouse with SNPs]

To further investigate the genetic mechanism of the mutant mice(B6-Co) with hereditary corneal opacity phenotype obtained by ENU-induced mutagenesis from B6 in previous study, SNP markers were used to map the mutant gene of B6-Co mice. F2 generation mice were bred by backcrossing (B6-CoPxD2 )F1 with D2 and the DNA samples of F2 mutant mice were extracted from the tails. Five SNP sites that showed differences between B6 and D2 strains nearby the located region on chromosome 13 were screened from MGI database. Five SNPs, PCR-RFLP and linkage analyses were carried out to map the mutant gene. The result showed that the mutant gene was located between 112 546 283~113 397 654 bp on chromosome 13. There are five identified genes including Map3k1 that is associated with eye morphogenesis and eyelid closure of mouse in this region. This suggests that Map3k1 is the most probable candidate mutant gene of B6-Co mice.