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[Cloning and expression of the mouse Doc-1R gene]
Xiu-jie Sheng1, Wei-qiang Zhou, Li Jiang
1Department of Laboratory Animal, China Medical University, Shenyang 110001, R. P. China.
Background & Objective:
Doc-1R gene is a new gene which was cloned in 1999. Recent studies suggest that Doc-1R gene is a potential tumor suppressor gene. In order to study the function of the Doc-1R gene, The aim of this study was to obtain its genomic sequences and to analyze its expression in the tissues.
Method:
According to the cDNA sequences of Doc-1R gene, the gene-specific primers were designed and synthesized. The sequence of Doc-1R gene was cloned through nested PCR using the Genome Walker kit. The sequence and splice donor/acceptor site were analyzed. Using the reverse transcription polymerase chain reaction(RT-PCR), the expression of Doc-1R gene in thirteen tissues, including mouse liver, spleen, pancreas, kidney, lung, intestine, heart, brain, bone, muscle, bladder, ovary, spermary were determined.
Results:
The mouse Doc-1R gene has been obtained by two times genomic walking. This gene spans 2,787 bp and contains four exons and three introns. All of the splice donor/acceptor site sequences were in accordance with the consensus "GT-AG" rule. RT-PCR experiments demonstrated Doc-1R gene was expressed in the 13 tissue samples.
Conclusion:
The authors have successfully cloned the mouse Doc-1R gene, which will be the foundation for further investigating the function of that gene. The expression pattern suggests that the Doc-1R gene is a housekeeping gene which is important to keep the function of tissues and organs.