Title

甲醇畢赤酵母Pichia methanolica的高效電轉化方法

Translated Titles

Electrotransformation of Pichia methanolica

Authors

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)

Key Words

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

PublicationName

食品與發酵工業

Volume or Term/Year and Month of Publication

30卷5期(2004 / 05 / 31)

Page #

35 - 39

Content Language

簡體中文

Chinese Abstract

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。

English Abstract

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.

Topic Category 工程學 > 化學工業