Electrotransformation of Thermoanaerobacter ethanolicus JW200

Authors: Peng, Hui; Fu, Bailing; Mao, Zhonggui; Shao, Weilan

Source: Biotechnology Letters, Volume 28, Number 23, December 2006 , pp. 1913-1917(5)

Publisher: Springer

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Abstract:

Electrotransformation of Thermoanaerobacter ethanolicus JW200 was achieved using the plasmid, pTE16, and a pUC-based suicide vector, pTEA2. The construct pTE16 is based on the Escherichia coli-Clostridium perfringens shuttle vector pJIR715 and contains a thermostable chloramphenicol (Cm) resistance cassette. Evidence supporting transformation was provided by extracting plasmid pTE16 from presumptive transformants of T. ethanolicus and by PCR specific to the chloramphenicol acetyltransferase (cat) gene on the vector pTEA2. Transformation frequencies of plasmid pTE16 and pTEA2 were 50 ± 7.4 and 30 ± 4.2 transformants per μg plasmid DNA. The results provide the first unequivocal gene transfer method functional in T. ethanolicus.

Keywords: electrotransformation; protoplast; shuttle vector; Thermoanaerobacter ethanolicus

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s10529-006-9184-6

Affiliations: 1: Email: wlshao@jsmail.com.cn

Publication date: 2006-12-01