Abstract: Objective To find mutational site of Ankrd55 and Ddx4 gene, clone and sequence these gene in B6-Co mutant strain mouse with cornea opacity phenotype. Methods The primers were designed according to mRNA. The target gene was amplified by PCR and RT-PCR in which the PCR templates were mRNA, and then recombinant vectors were transformed to competent cells after DNA fragments were connected to the T vector. The positive clones were selected and plasmid DNA extraction was detected by electrophoresis. The fragments by EcoRI digestion for identification of positive clones and at the end sequencing. Results The C base in sequence of Ddx4 chromosome 13 113 412 349 was replaced by A by sequence alignment with genome data base, with coding protein Pro replacement by Gln. Conclusion There is a single base mutation in Ddx4 gene, and the mutation may relate to the EOB phenotype of B6-Co mice. And it needs to be studied further.

Key words: B6 - Co mice, Corneal turbidity, PCR amplification, Gene clone