嗜杀苹果酒酵母的构建

The Construction of Killer Cider Yeast

采用核融合缺陷原生质体融合技术 ,将嗜杀质粒供体菌 5 0 45 (α ,his4,kar1— 1[KIL—K1 ]rho+ )中的嗜杀质粒转移到受体菌苹果酒酵母AW中去。用 3 5 %PEG ,3 0℃诱导融合 2 0min ,经嗜杀活性检出、小型酿造实验、遗传稳定性实验、细胞大小及体积的测定、dsRNA质粒的提取及琼脂糖凝胶电泳等实验 ,筛选出 4株遗传性状稳定、具有嗜杀活性且发酵性能良好的融合子F12、F3 0、F5 7、F65。

Killer plasmid was introduced into cider yeast by using protoplast fusion technique The donor strain 5045(α,his4,kar1 1 rho +) was karyogamy defective Two kinds of protoplast were impelled to fuse for 20min at 30℃ in the presence of PEG-6000 After killer character selecting, brewing in laboratory scale, generating more than ten times on selective medium, studying on cell morphology, preparing of dsRNA killer plasmid, agarose gel electrophoresis and so on, four fustants F12, F30, F57, F65 were obtained The results revealed that they were stable heteroplasmons with killer activity and good brewing characters