Bovin CMYA1 muscle specific expression core promoter and preparation method thereof

Abstract

The invention discloses a bovin CMYA1 muscle specific expression core promoter and a preparation method thereof. The preparation method comprises the following steps: firstly cloning a full-length fragment 1155bp of the bovin CMYA1 muscle specific expression core promoter, building a pEGFP-1-CMYA1 promoter carrier, and transfecting mice C2C12 myoblast, bovine skeletal muscle satellite cells and bovine fibroblast to prove the muscle specificity expression of the carrier; then, carrying out the deficiency research on CMYA1 promoter, totally designing 8 deletion fragments of the CMYA1 promoter, respectively building deletion fragment carriers of pGL3-basic-CMYA1 promoters, and transfecting C2C12 cells; and finally verifying, thereby obtaining the CMYA1 core promoter of which the gene sequence is SQ2. The CMYA1 promoter is specifically expressed in muscle cells, and the promotion activity is high; moreover, such the characteristic can be utilized by people to make a research on the development and differentiation of muscles.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering in the field of biotechnology, in particular to a bovine CMYA1 muscle-specific expression core promoter and a preparation method. Background technique [0002] 1. Research progress of CMYA1 gene. Different species of CMYAl (Cardiomyopathy associated protein 1, primary cardiomyopathy-related protein 1) contain five highly conserved domains, which are proline-rich regions and 16 amino acid repeats sequence (also called Xin repeat sequence), DNA binding domain, SH3 structural motif and nuclear localization signal. The bovine CMYA1 gene (CMYA1Gene ID: 509670) is located at Chromosome 22: 12549676~12558895bp, has two exons and one intron, encodes 1820 amino acids, and the total length of the coding region is 6272bp. The deduced protein isoelectric point (Isoelectric point) : 6.4162, molecular weight (Molecular weight): 194814.91g / mol. [0003] The CMYA1 gene was first discovered in the at...

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Problems solved by technology

[0005] Some researchers in China have also isolated and cloned the promoters of some genes, such as the cloning and verification of the mouse MCK promoter. The mouse MCK promoter was cloned to 1355bp, and the core promoter fragment was not screened for deletion. Some researchers also cloned the goat The MyoG gene promoter is 1200bp, and the promoter deletion study has not been carried out, and the core promoter fragment is obtained

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