A novel arabinose-inducible genetic operation system developed for Clostridium cellulolyticum

Biotechnology for Biofuels
Jie ZhangQiu Cui

Abstract

Clostridium cellulolyticum and other cellulolytic Clostridium strains are natural producers of lignocellulosic biofuels and chemicals via the consolidated bioprocessing (CBP) route, and systems metabolic engineering is indispensable to meet the cost-efficient demands of industry. Several genetic tools have been developed for Clostridium strains, and an efficient and stringent inducible genetic operation system is still required for the precise regulation of the target gene function. Here, we provide a stringent arabinose-inducible genetic operation (ARAi) system for C. cellulolyticum, including an effective gene expression platform with an oxygen-independent fluorescent reporter, a sensitive MazF-based counterselection genetic marker, and a precise gene knock-out method based on an inducible ClosTron system. A novel arabinose-inducible promoter derived from Clostridium acetobutylicum is employed in the ARAi system to control the expression of the target gene, and the gene expression can be up-regulated over 800-fold with highly induced stringency. The inducible ClosTron method of the ARAi system decreases the off-target frequency from 100% to 0, which shows the precise gene targeting in C. cellulolyticum. The inducible effect o...Continue Reading

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Citations

May 23, 2015·Applied Microbiology and Biotechnology·Elizabeth B MearlsLee R Lynd
Dec 18, 2015·Chembiochem : a European Journal of Chemical Biology·Dennis BinderThomas Drepper
Aug 28, 2020·Journal of Industrial Microbiology & Biotechnology·SongI HanGeorge N Bennett
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Feb 28, 2018·Frontiers in Microbiology·Rochelle C JosephNicholas R Sandoval
Aug 7, 2018·Journal of Biotechnology·Jie ZhangYi Wang

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Methods Mentioned

BETA
gene knock-out
PCR

Software Mentioned

ClosTron
ARAi

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