| 甲醇毕赤酵母Pichia methanolica的高效电转化方法 |
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| 引用本文： | 孙亚范,王海宽,向微,杜连祥,路福平. 甲醇毕赤酵母Pichia methanolica的高效电转化方法[J]. 食品与发酵工业, 2004, 30(5): 35-39 |
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| 作者姓名： | 孙亚范 王海宽 向微 杜连祥 路福平 |
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| 作者单位： | 天津科技大学食品科学与生物工程学院,天津,300222 |
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| 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。
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| 关 键 词： | 甲醇毕赤酵母 高效电转化 转化率 |
| 修稿时间： | 2003-11-27 |
Electrotransformation of Pichia methanolica |
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| Sun Yafan,Wang Haikuan,Xiang Wei,Du Lianxiang,Lu Fuping. Electrotransformation of Pichia methanolica[J]. Food and Fermentation Industries, 2004, 30(5): 35-39 |
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| Authors: | Sun Yafan Wang Haikuan Xiang Wei Du Lianxiang Lu Fuping |
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| Affiliation: | Sun Yafan,Wang Haikuan,Xiang Wei,Du Lianxiang,Lu Fuping Department of Food Science and Biology Engineering,Science and Technology University,Tianjin,300222 |
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| Abstract: | 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 forPichia methanolica was at intermediate logarithmic phage. Under the conditions of voltage of 600 Vand 30 μg/mL plasmid DNA and 0.2cm cuvettes, the transformation efficiency reach its maximum at57 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 ashost cells. |
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| Keywords: | Pichia methanolica efficient elect rotransformation transformation efficiency |
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