Cloning and expression of GLUT4 cDNA in E.coli / 西安交通大学学报(医学版)

ABSTRACT

Objective To acquire glucose transporter 4 (GLUT4) cDNA from tissues of human muscle by RT-PCR, and to clone and express GLUT4 cDNA in E.coli. Methods GLUT4 cDNA primers were designed first and ascertained by detecting through Genbank on NCBI and DNA Star, then by using specific primers, the subjective cDNA segment was acquired by RT-PCR from human tissue. After that, it was cloned into cloning vector of pGEM-3zf(-) and sequenced automatically. Finally the subjective cDNA was cloned into vector of pBV220 and expressed in E.coli. Results The desired DNA could be acquired from tissues of human muscle by RT-PCR using special primers, the acquired cDNA fragment could be cloned into vector of pGEM-3zf(-) and sequenced. The GLUT4 cDNA sequence was highly conserved. GLUT4 cDNA could be expressed in E.coli, too. Conclusion The entire GLUT4 cDNA can be acquired from human muscular tissue by RT-PCR, and it can be expressed in E.coli.