光滑球拟酵母金属硫蛋白基因的亚克隆和表达研究

用BamHI和PstI双酶切质粒pMTIIa－91s，获得光滑球拟酵母金属硫蛋白基因（MTIIa）片段，将其亚克隆到M13载体，扩增并回收MTIIa基因片段，经Southern杂交验证插入片段正确后，分别将MTIIa基因片段插入到大肠杆菌表达载体pBV220和大肠杆菌一酵母菌穿梭载体YIp352中，转化大肠杆菌DH5a，使其对CUSO₄的抗性提高0．7mol／L左右；转化酿酒酵母受体菌YS59，使YS59对CUSO4的抗性提高1．5mol／L左右。结果证明光滑球拟酵母MTIIa基因在

THE SUBCLONING AND EXPRESSION OF THE METALLOTHIONEIN-IIa GENE OF CANDIDA GLABRATA

The MTIIa gene of Candida glabrata was gained from plasmid pMTIIa--93s digested with restrichon enzymes BamHI and PstI, then subcloning it into M13 vector. It was confirmed that the insert fadment was right by Southern blotting. Proliferating MTIIa gene and ligating it into E. coli expression vector pBV220 and E.coli-yeast shuttle vector Yip352. The ligation mixtues were transformed into E. coli DH5 and S. cerevisiae YS59 respectively. The experiment of resistance to CuSO4. proved that the DH5 cells with the recombinant plasmids were 0.7mol / L higher than the cells without them, and the YS59 cells with the recombinant shuttle vectors were 1.5mol/L higher than the cells without them. According to these tesults, it proved that the MTIIa gene of Candida glabrata was expressed efficienUy.