| Products/Services Used | Details | Operation |
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| Gene Synthesis | A markerless deletion of the coxL gene (MSMEG_0746) was constructed by allelic exchange mutagenesis. Briefly, a 2245 bp fragment containing the fused left and right flanks of the MSMEG_0746 gene was synthesized by GenScript. This fragment was cloned into the SpeI site of the mycobacterial shuttle plasmid pX33 [68] with E. coliTOP10 and transformed into M. smegmatis mc2155 electrocompetent cells. To allow for temperature-sensitive vector replication, the transformants were incubated on LBT-gentamycin agar at 28°C for five days until colonies were visible. Catechol-reactive colonies were sub-cultured on to LBT-gentamycin agar plates incubated at 40°C for three days to facilitate the first recombination of the coxL flanks into the chromosome. | Get A Quote |
Carbon monoxide (CO) is a ubiquitous atmospheric trace gas produced by natural and anthropogenic sources. Some aerobic bacteria can oxidize atmospheric CO and, collectively, they account for the net loss of ~250 teragrams of CO from the atmosphere each year. However, the physiological role, genetic basis, and ecological distribution of this process remain incompletely resolved. In this work, we addressed these knowledge gaps through culture-based and culture-independent work. We confirmed through shotgun proteomic and transcriptional analysis that the genetically tractable aerobic soil actinobacterium Mycobacterium smegmatis upregulates expression of a carbon monoxide dehydrogenase by 50-fold when exhausted f... More
