[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.


CONCLUSIONS
The genomic walking strategy is simple, efficient and reliable; it is an ideal method of cloning genomic fragments.
Sheng
X
X
Department of Laboratory Animal, China Medical University, Shenyang, Liaoning 110001 P.R.China. Xjsheng@hotmail.com
Jiang
L
L
Zhou
W
W
Wang
T
T
Zhang
X
X
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D005091:Exons; D016147:Genes, Tumor Suppressor; D016678:Genome; D006801:Humans; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018076:DNA, Complementary; C485718:Doc-1r protein, mouse; D015513:Oncogene Proteins