7 research outputs found
Site-Specific Covalent Labeling of RNA by Enzymatic Transglycosylation
We demonstrate the site-specific
incorporation of nucleoÂbase
derivatives bearing fluoroÂphores or affinity labels into a short
RNA stem loop recognition motif by exchange of a guanine residue.
The RNA-TAG (transÂglycosylÂation at guanosine) is carried
out by a bacterial (<i>E. coli</i>) tRNA guanine transÂglycosylÂase
(TGT), whose natural substrate is the nitrogenous base PreQ<sub>1</sub>. Remarkably, we have successfully incorporated large functional
groups including biotin, BODIPY, thiazole orange, and Cy7 through
a polyÂethylene glycol linker attached to the exocyclic amine
of PreQ<sub>1</sub>. Larger RNAs, such as mRNA transcripts, can be
site-specifically labeled if they possess the 17-nucleotide hairpin
recognition motif. The RNA-TAG methodology could facilitate the detection
and manipulation of RNA molecules by enabling the direct incorporation
of functional artificial nucleoÂbases using a simple hairpin
recognition element
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Expanding the diversity of mycobacteriophages: insights into genome architecture and evolution.
Mycobacteriophages are viruses that infect mycobacterial hosts such as Mycobacterium smegmatis and Mycobacterium tuberculosis. All mycobacteriophages characterized to date are dsDNA tailed phages, and have either siphoviral or myoviral morphotypes. However, their genetic diversity is considerable, and although sixty-two genomes have been sequenced and comparatively analyzed, these likely represent only a small portion of the diversity of the mycobacteriophage population at large. Here we report the isolation, sequencing and comparative genomic analysis of 18 new mycobacteriophages isolated from geographically distinct locations within the United States. Although no clear correlation between location and genome type can be discerned, these genomes expand our knowledge of mycobacteriophage diversity and enhance our understanding of the roles of mobile elements in viral evolution. Expansion of the number of mycobacteriophages grouped within Cluster A provides insights into the basis of immune specificity in these temperate phages, and we also describe a novel example of apparent immunity theft. The isolation and genomic analysis of bacteriophages by freshman college students provides an example of an authentic research experience for novice scientists
Expanding the diversity of mycobacteriophages: insights into genome architecture and evolution.
Mycobacteriophages are viruses that infect mycobacterial hosts such as Mycobacterium smegmatis and Mycobacterium tuberculosis. All mycobacteriophages characterized to date are dsDNA tailed phages, and have either siphoviral or myoviral morphotypes. However, their genetic diversity is considerable, and although sixty-two genomes have been sequenced and comparatively analyzed, these likely represent only a small portion of the diversity of the mycobacteriophage population at large. Here we report the isolation, sequencing and comparative genomic analysis of 18 new mycobacteriophages isolated from geographically distinct locations within the United States. Although no clear correlation between location and genome type can be discerned, these genomes expand our knowledge of mycobacteriophage diversity and enhance our understanding of the roles of mobile elements in viral evolution. Expansion of the number of mycobacteriophages grouped within Cluster A provides insights into the basis of immune specificity in these temperate phages, and we also describe a novel example of apparent immunity theft. The isolation and genomic analysis of bacteriophages by freshman college students provides an example of an authentic research experience for novice scientists