Article
An upp-based markerless gene replacement method for genome reduction and metabolic pathway engineering in Pseudomonas mendocina NK-01 and Pseudomonas putida KT2440.
Journal of microbiological methods - 1 Jun 2015
Wang Yuanyuan, Zhang Chi, Gong Ting, Zuo Zhenqiang, Zhao Fengjie, Fan Xu, Yang Chao, Song Cunjiang
Abstract excerpt
A markerless gene replacement method was adapted by combining a suicide plasmid, pEX18Tc, with a counterselectable marker, the upp gene encoding uracil phosphoribosyltransferase (UPRTase), for the medium-chain length polyhydroxyalkanoates (PHA(MCL))-producing strain Pseudomonas mendocina NK-01. An NK-01 5-fluorouracil (5-FU) resistant background strain was first constructed by deleting the chromosomal upp gene....
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