甲醇毕赤酵母Pichia methanolica的高效电转化方法  

Electrotransformation of Pichia methanolica

作　　者：孙亚范[1] 王海宽[1] 向微[1] 杜连祥[1] 路福平[1] 

机构地区：[1]天津科技大学食品科学与生物工程学院

出　　处：《食品与发酵工业》2004年第5期35-39,共5页Food and Fermentation Industries

摘　　要：采用电穿孔的方法对甲醇毕赤酵母进行外源DNA的转化。通过调整参数,对影响电转化效率的主要因素进行探索,建立了质粒pMETA-Lip对甲醇毕赤酵母PMAD16的高效电转化方法。结果表明,当采用对数生长中期的菌体制备感受态细胞、质粒DNA浓度为30μg/mL、采用0.2cm电转杯、电压为600 V时,转化率达到最大值,为57个转化子/μg质粒DNA。经抽样鉴定所得到的转化子均为阳性克隆。为外源基因在甲醇毕赤酵母中的表达奠定了基础。Electrotransformation, a highly efficient and easy to apply technique, is used to intro- duce plasmid pMETA-Lip into Pichia methanolica in this work. The optimal electroporated stage for Pichia methanolica was at intermediate logarithmic phage. Under the conditions of voltage of 600 V and 30 μg/mL plasmid DNA and 0.2cm cuvettes, the transformation efficiency reach its maximum at 57 transformants/μg plasmid DNA. The results showed that all of transformants were positive clones. The experiments provided a model for highly efficient electrotransformation of Pichia methanolica as host cells.

关 键 词：甲醇毕赤酵母 高效电转化方法 电穿孔方法 外源DNA 

分 类 号：TQ920[轻工技术与工程—发酵工程]