Cloning of glycophorin A cDNA and construction of its expression plasmid for yeast two hybrid system / 中国病理生理杂志

ABSTRACT

AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.