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利用nah-lux转录融合通过生物发光监测萘的分解代谢。

Monitoring of naphthalene catabolism by bioluminescence with nah-lux transcriptional fusions.

作者信息

Burlage R S, Sayler G S, Larimer F

机构信息

Center for Environmental Biotechnology, University of Tennessee, Knoxville 37996.

出版信息

J Bacteriol. 1990 Sep;172(9):4749-57. doi: 10.1128/jb.172.9.4749-4757.1990.

Abstract

We have demonstrated the efficacy of a light-generating genetic construction in describing the induction of a nah operon for the catabolism of naphthalene. A fragment from plasmid NAH7, which contains the promoter for the upper pathway of degradation, was transcriptionally fused to the lux genes of Vibrio fischeri. A Pseudomonas strain containing this construction is inducible to high levels of light production in the presence of a suitable substrate and the nahR regulatory gene product. This system was used to examine catabolic activity in a unique manner under a variety of growth conditions. Induction of bioluminescence was demonstrated to coincide with naphthalene degradation in all cases through the use of mineralization assays. A significant delay in bioluminescence and biodegradation was observed when naphthalene was added to batch cultures that were growing exponentially. These results suggest that the metabolism of naphthalene by this Pseudomonas strain is optimal when the growth rate of the culture is slow and is greatly reduced during exponential growth.

摘要

我们已经证明了一种发光基因构建体在描述萘分解代谢中nah操纵子的诱导方面的功效。来自质粒NAH7的一个片段,其包含降解上途径的启动子,被转录融合到费氏弧菌的lux基因上。含有这种构建体的假单胞菌菌株在合适的底物和nahR调节基因产物存在的情况下可诱导产生高水平的光。该系统被用于以独特的方式在各种生长条件下检测分解代谢活性。通过使用矿化分析表明,在所有情况下生物发光的诱导都与萘的降解同时发生。当将萘添加到指数生长的分批培养物中时,观察到生物发光和生物降解有显著延迟。这些结果表明,当培养物的生长速率缓慢时,这种假单胞菌菌株对萘的代谢是最佳的,而在指数生长期间会大大降低。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/678e/213127/4e36beddbb31/jbacter00123-0022-a.jpg

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