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Detection and quantification of glycosylated queuosine modified tRNAs by acid denaturing and APB gels

RNA. 2020; 
Wen Zhang, Ruyi Xu, Żaneta Matuszek, Zhen Cai, Tao Pan
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Molecular Biology Reagents … application of standard denaturing gels supplemented with a boric acid derivative (Igloi and … the cis-diol and boric acid derivatives in the gel, so a limitation of the method is the difficulty … incubated with streptavidin-HRP conjugate (Genscript, M00091) in 30 ml hybridization buffer … Get A Quote

摘要

Queuosine (Q) is a conserved tRNA modification in bacteria and eukaryotes. Eukaryotic Q-tRNA modification occurs through replacing the guanine base with the scavenged metabolite queuine at the wobble position of tRNAs with UN anticodon (Tyr, His, Asn, Asp) by the QTRT1/QTRT2 heterodimeric enzyme encoded in the genome. In humans, Q-modification in tRNA and tRNA are further glycosylated with galactose and mannose, respectively. Although galactosyl-Q (galQ) and mannosyl-Q (manQ) can be measured by LC/MS approaches, the difficulty of detecting and quantifying these modifications with low sample inputs has hindered their biological investigations. Here we describe a simple acid denaturing gel and nonradioactive nort... More

关键词

acid denaturing PAGE, galactosyl-queuosine, mannosyl-queuosine, queuosine