Functional Role of Methylation of G518 of the 16S rRNA 530 Loop by GidB in Mycobacterium tuberculosis (original) (raw)

“…Automated annotation was conducted using RoboOligo [27] and in-house software. The criterion for detection of a particular RNase digestion product based on MS/MS analysis was at least 80% of the expected c- and y-type ions [28] were found during spectral annotation, except for 7-methylguanosine containing digestion products, which fragment uniquely [29]. …”

Section: Methodsmentioning

“…The studies by Wong et al showed that methylation by GidB significantly enhanced streptomycin binding to the ribosome, but some clinical isolates with gidB mutations were also found to be streptomycin sensitive [23,85]. Further studies suggest particular point mutations in gidB are responsible for streptomycin resistance [57,87,90].…”

Section: Antibiotic Resistancementioning

“…The RNA digestion products were resolved on an XBridge C18 column (1.0 Â 150 mm; Waters, Milford, MA) by a 70-minute gradient elution with mobile phase C and mobile phase D (50% mobile phase C, 50% methanol) at a flow rate of 30 ml/min using a Thermo Surveyor highperformance liquid chromatography system and a Thermo Micro AS autosampler. The tuning and MS methods were essentially the same as previously described (Wong et al, 2013). Mass values were restricted to a scan range of m/z 600-2000, and data-dependent collision-induced dissociation MS/MS was performed for the top four most abundant ions before keeping them in the exclusion list for 30 seconds.…”

Section: Methodsmentioning