A Method for Determining the Core Promoter of Guanling Cattle Myodi Gene Using Dual Luciferase Reporter Genes

Abstract

The invention discloses a method of using dual luciferase reporter genes to determine the MyoD Ⅰ The method for the gene core promoter comprises the following steps: constructing a recombinant vector containing the full-length fragment of the Guanling cattle MyoDI gene promoter and a subcloned fragment; cultivating and plating cells related to skeletal muscle growth; preparing the promoter recombinant sub / lipid The mixture of plastids was used to transfect the cells obtained in step 2); the transfected cells obtained in step 3) were detected for dual luciferase activity by a dual luciferase reporter gene. The present invention has the advantages of high sensitivity, fast detection speed, low cost, and no need to use radioactive isotopes. The pGL3-Basic reporter gene used can detect the insertion through the expression of the luciferase reporter gene because it has no promoter and enhancer. Promoter activity, and compare the strength of the promoter, and then preliminarily determine the core promoter region.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a method of using dual luciferase reporter genes to determine the MyoDI Methods for gene core promoters. Background technique [0002] MyoDI Its full name is myogenic differentiation Ⅰ, and its Chinese name is myogenic determinant factor. It is an important positive regulatory gene in the process of skeletal muscle growth and development. In the regulation of myogenic muscle-specific gene transcription, MyoDI Acting as a master switch, it can bind to the promoter regions of myogenin, creatine kinase CK, myosin, desmin and other genes to play an important regulatory role and promote their transcriptional activation. MyoDI As the best model to study cell differentiation, the mediated muscle differentiation mechanism has become the focus of many scholars' research and is particularly important. At present, there are many related studies on the expression and regulation of this g...

AI Technical Summary

Problems solved by technology

At present, there are many related studies on the expression and regulation of this gene in the myocyte formation mechanism of mice, chickens and pigs at home and abroad. There have been some studies on cattle, including genetic variation and function studies, but mainly focused on post-transcriptional and translational levels, MyoDI The molecular mechanism of gene transcription regulation is still unclear

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