Cloning of glycophorin A cDNA and construction of its expression plasmid for yeast two hybrid system
- VernacularTitle:酵母双杂合系统BD端血型糖蛋白A表达质粒的构建
- Author:
Hongtao LI
;
Guohui FU
;
Xiaoshu JIANG
;
Baoshan ZHANG
;
Xiangan KONG
- Publication Type:Journal Article
- Keywords:
Glycophorin;
Yeasts;
Membrane proteins
- From:
Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
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.
