关岭牛MyoD I基因启动子真核表达载体的构建及其在小鼠C2C12细胞中的表达

doi:10.11843/j.issn.0366-6964.2015.05.006

Abstract

Abstract:

The aim of this study was to analyze promoter activity of 4 different lengths MyoD I promoters in Guanling cattle and determine preliminarily the position of the core gene promoter.The relative expression quantity was detected in different tissues of Guanling cattle by real-time quantitative PCR.The DNA extracted from Guanling cattle blood was as a template and the phosphorylated 5'end PCR primers was designed to amplify 4 different lengths MyoD I promoters in Guanling cattle，ultimately accurately replaced the CMV region of pEGFP-N3 vector and got eukaryotic expression vector pEGFP-N3-MyoD I.Then the recombinant plasmid was transiently transfected into mouse C2C12 cells by liposome method.The aim was to verify promoter activity of MyoD I promoter by Luminous intensity of mouse C2C12 cells.The expression level of MyoD I gene was the highest in muscle，the fragment was inserted into an expression vector，the recombinant plasmid could be successfully expressed in mouse C2C12 cells with green fluorescence under the excitation light.In this study,eukaryotic expression vector pEGFP-N3-MyoD I is successfully constructed，4 promoters constructed show promoter activity，expression quantity of P₃ promoter is the highest，so P₃ promoter is regarded as the core promoter of MyoD I gene.

CLC Number:

S823
S813.3

HUAN Cong-cong，XU Hou-qiang，CHEN Wei，CHEN Xiang，ZHAO Jia-fu，ZHANG Wen，ZHOU Di，XIA Dan. Construction of MyoD I Promoter Eukaryotic Expression Vector in Guanling Cattle and Its Expression in Mouse C2C12 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(5): 719-727.

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Construction of MyoD I Promoter Eukaryotic Expression Vector in Guanling Cattle and Its Expression in Mouse C2C12 Cells

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