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作为环境生物传感器的重组微生物:通过携带fabA'::lux融合基因的大肠杆菌检测污染物

Recombinant microorganisms as environmental biosensors: pollutants detection by Escherichia coli bearing fabA'::lux fusions.

作者信息

Bechor Ofra, Smulski Dana R, Van Dyk Tina K, LaRossa Robert A, Belkin Shimshon

机构信息

Division of Environmental Sciences, The Fredy and Nadine Herrmann Graduate School of Applied Science, The Hebrew University, Jerusalem 91904, Israel.

出版信息

J Biotechnol. 2002 Mar 14;94(1):125-32. doi: 10.1016/s0168-1656(01)00423-0.

Abstract

A set of genetically engineered Escherichia coli strains was constructed, in which the promoter of the fabA gene is fused to Vibrio fischeri luxCDABE either in a multi-copy plasmid or as a single copy chromosomal integration. The fabA gene codes for beta-hydroxydecanoyl-ACP dehydrase, a key enzyme in the synthesis of unsaturated fatty acids, and is induced when fatty acid biosynthesis pathways are interrupted. A dose-dependent and highly sensitive bioluminescent response to a variety of chemicals was controlled by the fadR gene. A tolC mutant E. coli host displayed generally lower detection threshold for toxicants. A chromosomal integration of a single copy of the fabA'::lux fusion led to a markedly lower background luminescence, but did not yield an improvement in overall performance. It is proposed that these or similarly constructed reporters of fatty acid biosynthesis inhibition may serve as novel microbial toxicity biosensors.

摘要

构建了一组基因工程大肠杆菌菌株,其中fabA基因的启动子与费氏弧菌luxCDABE融合,融合方式为多拷贝质粒或单拷贝染色体整合。fabA基因编码β-羟基癸酰-ACP脱水酶,这是不饱和脂肪酸合成中的关键酶,当脂肪酸生物合成途径中断时会被诱导。对多种化学物质的剂量依赖性和高灵敏度生物发光反应由fadR基因控制。tolC突变的大肠杆菌宿主对毒物的检测阈值通常较低。fabA'::lux融合单拷贝的染色体整合导致背景发光明显降低,但整体性能并未得到改善。有人提出,这些或类似构建的脂肪酸生物合成抑制报告基因可作为新型微生物毒性生物传感器。

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