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Nanaerobic growth enables direct visualization of dynamic cellular processes in human gut symbionts

Proc Natl Acad Sci U S A. 2020; 
Leonor García-Bayona, Michael J Coyne, Noam Hantman, Paula Montero-Llopis, Salena S Von, Takeshi Ito, Michael H Malamy, Marek Basler, Blanca Barquera, Laurie E Comstock
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Gene Synthesis … The genes for the red fluorescent proteins, DsRed2, mKate2, and TagRFP were codon optimized for Bacteroides (Fig S3) and synthesized by Genscript. Each gene was amplified with the primers listed in Table S4 and used to replace the super folder GFP gene and the … Get A Quote

摘要

Mechanistic studies of anaerobic gut bacteria have been hindered by the lack of a fluorescent protein system to track and visualize proteins and dynamic cellular processes in actively growing bacteria. Although underappreciated, many gut "anaerobes" are able to respire using oxygen as the terminal electron acceptor. The oxygen continually released from gut epithelial cells creates an oxygen gradient from the mucus layer to the anaerobic lumen [L. Albenberg et al., Gastroenterology 147, 1055-1063.e8 (2014)], with oxygen available to bacteria growing at the mucus layer. Here, we show that species are metabolically and energetically robust and do not mount stress responses in the presence of 0.10 to 0.14% oxygen,... More

关键词

Bacteroides, GFP, fluorescent proteins, microbiota, nanaerobic
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