[Cloning of the mouse Doc-1R gene by genomic walking].

OBJECTIVE

To obtain the genomic sequences of the mouse Doc-1R gene.

METHODS

Gene-specific primers were designed and synthesized based on the cDNA sequences of the mouse Doc-1R gene. With the use of genomic walking strategy, the mouse genomic walking library was amplified by the polymerase chain reaction(PCR). Mouse genomic library constructed with a special adaptor was utilized as a template to amplify the desired fragment by nested PCR.

RESULTS

A desired fragment of 1.5 kb was obtained. Sequence analysis of the desired fragment confirmed that the genomic cloning of the Doc-1R gene was successful. This gene contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 'GT-AG' rule.

CONCLUSION

The genomic walking strategy is simple, efficient and reliable; it is an ideal method of cloning genomic fragments.