Campen R K, Duester G L, Holmes W M, Young J M
J Bacteriol. 1980 Dec;144(3):1083-93. doi: 10.1128/jb.144.3.1083-1093.1980.
The arrangement of transfer ribonucleic acid (RNA) genes in the chromosome of Escherichia coli K-12 (C600) was examined with the techniques of restriction endonuclease digestion and Southern blotting. The number and size of restriction fragments containing transfer or ribosomal RNA sequences or both were estimated by a variety of restriction endonucleases, including EcoRI, BglI, SmaI, SalI, BamHI, and PstI. EcoRI liberated a minimum of 27 fragments which hybridized to transfer RNA and 16 which hybridized to ribosomal RNA. Enzymes which did not cut within the ribosomal RNA operons (PstI and BamHI) liberated 16 and 13 fragments, respectively, which hybridized to transfer RNA. Five PstI and six BamHi fragments also hybridized to ribosomal RNA, suggesting that there may be at least 11 chromosomal locations distinct from ribosomal RNA operons which encode transfer RNA genes. In addition, our data indicated that several transfer RNA genes may be very close to the 5' proximal ends of certain ribosomal RNA operons and close to the 3' distal ends of all seven ribosomal RNA operons. Similar studies have been carried out with 22 purified species of transfer RNA, and we report here the number and size of EcoRI restriction fragments which hybridize to these transfer RNA species.
运用限制性内切酶消化和Southern印迹技术，对大肠杆菌K - 12（C600）染色体中转录核糖核酸（RNA）基因的排列进行了检测。通过多种限制性内切酶，包括EcoRI、BglI、SmaI、SalI、BamHI和PstI，对含有转运或核糖体RNA序列或两者兼有的限制性片段的数量和大小进行了估算。EcoRI释放出至少27个与转运RNA杂交的片段以及16个与核糖体RNA杂交的片段。在核糖体RNA操纵子内部不进行切割的酶（PstI和BamHI）分别释放出16个和13个与转运RNA杂交的片段。5个PstI片段和6个BamHI片段也与核糖体RNA杂交，这表明可能存在至少11个不同于核糖体RNA操纵子的染色体位置，这些位置编码转运RNA基因。此外，我们的数据表明，几个转运RNA基因可能非常靠近某些核糖体RNA操纵子的5'近端，并且靠近所有7个核糖体RNA操纵子的3'远端。我们对22种纯化的转运RNA物种进行了类似研究，在此报告与这些转运RNA物种杂交的EcoRI限制性片段的数量和大小。
