Transformation Conditions of Thermoanaerobacter ethanolicus JW200

China Biotechnology

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Transformation Conditions of Thermoanaerobacter ethanolicus JW200

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Abstract Abstract: It is necessary to develop a genetic transformation system for Thermoanaerobacter. ethanolicus JW200 to study gene function and regulation. To realize this, the gene transfer techniques for this organism were first developed in this paper. Firstly, transformation using polyethylene glycol (PEG6000) was obtained using T. ethanolicus-Escherichia coli shuttle plasmid pTE16. Secondly, electrotransformation was employed to introduce plasmid pTE16 into Thermoanaerobacter. ethanolicus. Transformation efficiency of PEG method was very low.，so electrotransformation procedures were optimized to enhance transformation frequency from 4±3.2 to 50±7.4 transformants per μg plasmid DNA. The most crucial factors conducive to achieve higher transformation efficiency included the appropriate degree of weaken cell wall and the chilling of cells during cell collection and washing. The results reported here provide the first unequivocal gene transfer method functional in T. ethanolicus that is suitable for molecular manipulations.

Key words： Optimization of condition Thermoanaerobacter ethanolicus transformation electrotransformation

Received: 06 July 2006 Published: 25 November 2006

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. Transformation Conditions of Thermoanaerobacter ethanolicus JW200. China Biotechnology, 2006, 26(11): 59-61.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I11/59

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