洋葱伯克霍尔德菌
生物
基因
载体(分子生物学)
发起人
洋葱伯克霍尔德菌复合体
遗传学
基因表达
表达式向量
分子生物学
伯克氏菌属
细菌
重组DNA
毒力
作者
Silvia T. Cardona,Miguel A. Valvano
出处
期刊:Plasmid
[Elsevier]
日期:2005-11-01
卷期号:54 (3): 219-228
被引量:140
标识
DOI:10.1016/j.plasmid.2005.03.004
摘要
Infection of the respiratory tract caused by Burkholderia cepacia complex poses a serious risk for cystic fibrosis (CF) patients due to the high morbidity and mortality associated with the chronic infection and the lack of efficacious antimicrobial treatments. A detailed understanding of the pathogenicity of B. cepacia complex infections is hampered in part by the limited availability of genetic tools and the inherent resistance of these isolates to the most common antibiotics used for genetic selection. In this study, we report the construction of an expression vector which uses the rhamnose-regulated PrhaB promoter of Escherichia coli. The functionality of the vector was assessed by expressing the enhanced green fluorescent protein (eGFP) gene (e-gfp) and determining the levels of fluorescence emission. These experiments demonstrated that PrhaB is responsive to low concentrations of rhamnose and it can be effectively repressed with 0.2% glucose. We also demonstrate that the tight regulation of gene expression by PrhaB promoter allows us to extend the capabilities of this vector to the identification of essential genes.
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