异源蛋白在酵母中的表达涉及到大量重组子的筛选鉴定.以直接煮沸2~5 min后的酵母菌落水悬浮液为模板,建立了酵母克隆的PCR快速筛选方法.介绍了PCR反应中减少引物二聚体、提高反应特异性及-80℃低温保存酿酒酵母感受态细胞等技巧.
Heterologous expression of recombinant proteins in yeast involves toilsome screening of a large number of recombinants from yeast cells. A quick and easy procedure was introduced for identifying recombinant clones from the yeast Saccharomyces cerevisiae by means of PCR without any prior DNA extraction and purification steps. This method involves a simple boiling step for 2~5 min of whole yeast cells suspended in water. Both the fresh and frozen competent S.cerevisiae cells can be used for transformation. The method of storing the competent yeast cells at -80℃ for future use is convenient and economic. A skill of decreasing the dimer products and increasing the specificity of PCR is recommended.
叶玲,刘建伟,刘静.酿酒酵母感受态细胞的低温保存及酵母菌落PCR-快速筛选鉴定[J].生物化学与生物物理进展,2003,30(6):956-959
复制生物化学与生物物理进展 ® 2025 版权所有 ICP:京ICP备05023138号-1 京公网安备 11010502031771号