title: Two-dimensional polyacrylamide-gel electrophoresis for purification of small RNAs specified by virulent coliphages T4, T5, T7 and BF23.
abstract: RNAs synthesized in Escherichia coli infected with virulent phages T4, T5, T7 and BF23 were labelled with 32PO4 3- after phage infection. [32P]RNAs of low molecular weight were separated by two-dimensional polyacrylamide gel electrophoresis, in which electrophoresis was carried out in two dimensions at different concentrations of acrylamide. The fractionated RNAs were characterized by RNA-fingerprint patterns made after T1 ribonuclease digestion. The two-dimensional gel of 10% yields 20% acrylamide was suitable for RNA of less than 200 nucleotides, while that of 5% yields 10% was preferred for RNAs of about 150--400 nucleotides. With T4 phage, 16 RNA species were separable on a single slab gel. Among those, 11 were identified as the known RNA species, including eight T4 tRNAs, one tRNA precursor and two non-tRNA molecules. In the case of T5 and BF23, more than 20 RNA species were separated on a slab gel; 15 or more RNAs were found in the 4-S RNA region, and several in 5-S and 6-S region. The RNA-fingerprint patterns of many BF23 RNAs were very similar to those of corresponding RNAs of T5. Pseudouridine and ribosylthymidine, minor nucleosides generally present in tRNA, were found in several BF23 4-S RNAs tested. Possibility of those BF23 4-S RNAs as tRNAs is discussed. With phage T7, three RNAs were detected, two of which were much smaller than tRNAs.
mesh_terms: D003090:Coliphages; D004267:DNA Viruses; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis*; D009841:Oligonucleotides / Q000032:analysis; D011560:Pseudouridine / Q000032:analysis; D012329:RNA, Bacterial / Q000032:analysis; D012343:RNA, Transfer / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis* / Q000096:biosynthesis; D012260:Ribonucleases; D012263:Ribonucleosides / Q000032:analysis; D012265:Ribonucleotides / Q000032:analysis; D013045:Species Specificity
chemical_list: D009841:Oligonucleotides; D012329:RNA, Bacterial; D012367:RNA, Viral; D012263:Ribonucleosides; D012265:Ribonucleotides; D011560:Pseudouridine; D012343:RNA, Transfer; D012260:Ribonucleases