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[Expression of human SRY gene and the DNA-binding property of its product]
1Department of Biochemistry and Immunology, Capital Institute of Pediatrics, Beijing 100020 P. R. China. cip@public.intercom.com.cn
Objective:
To investigate the role which human SRY gene plays in the regulation of the downstream gene.
Methods:
The fragment of SRY (sex determinating region on the Y chromosome) HMG domain was cloned into the expressing vector pET-15b. The hSRY gene recombinant plasmid-pETSY was transformed and expressed in E.coliBL21. The target protein was purified by pET His. Tag system. The DNA-binding retardation test and its competitive reaction were conducted between SRY protein and the fragment of Mullerian inhibiting substance (MIS) promoter.
Results:
The molecular weight of the expressed hSRY protein was shown to be approximately 21kD. The specific DNA binding property of SRY protein to the fragment of MIS was confirmed in the retardation test and its competitive reaction.
Conclusion:
The results suggest that the product of SRY gene can bind the MIS promoter region and may initiate the transcription of MIS.