甲醇畢赤酵母Pichia methanolica的高效電轉化方法

Electrotransformation of Pichia methanolica

孫亞範(Ya-Fan Sun)；王海寬(Hai-Kuan Wang)；向微(Wei Xiang)；杜連祥(Lian-Xiang Du)；路福平(Fu-Ping Lu)

甲醇畢赤酵母 ； 高效電轉化 ； 轉化率 ； Pichia methanolica ； efficient electrotransformation ； transformation efficiency

食品與發酵工業

30卷5期（2004 / 05 / 31）

35 - 39

簡體中文

採用電穿孔的方法對甲醇畢赤酵母進行外源DNA的轉化。通過調整參數，對影響電轉化效率的主要因素進行探索，建立了質粒pMETA-Lip對甲醇畢赤酵母PMAD16的高效電轉化方法。結果表明，當採用對數生長中期的菌體製備感受態細胞、質粒DNA濃度為30μg/mL、採用0.2cm電轉杯、電壓為600V時，轉化率達到最大值，為57個轉化子／μg質粒DNA。經抽樣鑒定所得到的轉化子均為陽性克隆。為外源基因在甲醇畢赤酵母中的表達奠定了基礎。

Electrotransformation, a highly efficient and easy to apply technique, is used to introduce 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 600V and 30μg/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.