کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1994385 | 1064775 | 2008 | 10 صفحه PDF | دانلود رایگان |
Here we describe the many applications of acid urea polyacrylamide gel electrophoresis (acid urea PAGE) followed by Northern blot analysis to studies of tRNAs and aminoacyl-tRNA synthetases. Acid urea PAGE allows the electrophoretic separation of different forms of a tRNA, discriminated by changes in bulk, charge, and/or conformation that are brought about by aminoacylation, formylation, or modification of a tRNA. Among the examples described are (i) analysis of the effect of mutations in the Escherichia coli initiator tRNA on its aminoacylation and formylation; (ii) evidence of orthogonality of suppressor tRNAs in mammalian cells and yeast; (iii) analysis of aminoacylation specificity of an archaeal prolyl-tRNA synthetase that can aminoacylate archaeal tRNAPro with cysteine, but does not aminoacylate archaeal tRNACys with cysteine; (iv) identification and characterization of the AUA-decoding minor tRNAIle in archaea; and (v) evidence that the archaeal minor tRNAIle contains a modified base in the wobble position different from lysidine found in the corresponding eubacterial tRNA.
Journal: Methods - Volume 44, Issue 2, February 2008, Pages 129–138