Construction of mouse MyoD vector and its expression in Escherichia coli / 实用口腔医学杂志

ABSTRACT

Objective:To identify the cDNA gene of mouse MyoD by restriction enzyme analysis, and to express the gene in Escherichia coli (E coli) using a protein expression vector.Methods: After the cDNA gene of mouse MyoD had been amplified and identified,it was inserted into expression vector pBV220 in which exogenous gene was controlled by R RP L promoters.The recombinant plasmid pBV-my was transformed into E coli DH5? and the bacteria were induced at 42 ℃ to express encoded protein.Results:The cDNA of mouse MyoD was sequenced correctly.When the engineered bacteria had been induced an anticipated 55 ku protein band from the bacteria was observed on SDS-PAGE gel and amounted to 30% of total bacterial protein. Conclusion:The cDNA of mouse MyoD has been successfully coloned and efficiently expressed in E coli.