relE作为阴性选择标记在致病性大肠杆菌中构建确定性基因构建体的应用及优化
Application and Optimization of relE as a Negative Selection Marker for Making Definitive Genetic Constructs in Uropathogenic Escherichia coli.
作者信息
Khetrapal Varnica, Mehershahi Kurosh S, Chen Siyi, Chen Swaine L
机构信息
Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, 1E Kent Ridge Road, NUHS Tower Block, Level 10, Singapore 119074, Singapore.
Genome Institute of Singapore, Infectious Diseases Group, 60 Biopolis street, Genome, #02-01, Singapore 138672, Singapore.
出版信息
Pathogens. 2016 Jan 18;5(1):9. doi: 10.3390/pathogens5010009.
Studies of Uropathogenic Escherichia coli (UPEC) pathogenesis have relied heavily on genetic manipulation to understand virulence factors. We applied a recently reported positive-negative selection system to create a series of unmarked, scarless FimH mutants that show identical phenotypes to previously reported marked FimH mutants; these are now improved versions useful for definitive assignment of phenotypes to FimH mutations. We also increased the efficiency of this system by designing new primer sites, which should further improve the efficiency and convenience of using negative selection in UTI89, other UPEC, and other Enterobacteriaceae.
对尿路致病性大肠杆菌（UPEC）发病机制的研究在很大程度上依赖于基因操作来了解毒力因子。我们应用了最近报道的正负选择系统来创建一系列无标记、无疤痕的FimH突变体，这些突变体表现出与先前报道的有标记FimH突变体相同的表型；它们现在是改进版本，有助于将表型明确归因于FimH突变。我们还通过设计新的引物位点提高了该系统的效率，这应该会进一步提高在UTI89、其他UPEC和其他肠杆菌科细菌中使用负选择的效率和便利性。
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