| abstract |
The method that the present invention provides the core promoter for determining miR 27a genes, 8 miR 27a promoter deletion fragments are expanded from mouse blood genomic DNA, structure restructuring Dual-Luciferase expression vector pGL3 D1 to pGL3 D8, the uciferase activity that 8 deletion fragments are detected after Chinese hamster ovary celI is transferred to, obtains core promoter area；This method identifies the core space of miR 27a, and new startup child resource is provided for genetic engineering and molecular breeding；The present invention also provides the method for transcription factor Myod binding sites in definite miR 27a gene core promoters, by overexpressing the Binding site for transcription factor in transcription factor Myod and rite-directed mutagenesis core promoter area, build saltant type fluorescent expression vector, uciferase activity is detected, determines Binding site for transcription factor；Apply to the research of domestic animal prolificacy molecular regulation mechanism for the expression regulation of miR 27a foundation is provided. |