Marconi R T, Hill W E
Department of Chemistry, University of Montana, Missoula 59812.
Nucleic Acids Res. 1988 Feb 25;16(4):1603-15. doi: 10.1093/nar/16.4.1603.
The accessibility of specific sequences in domain V of E. coli 23s rRNA in the 50S subunit to complementary oligodeoxyribonucleotides (cDNA) has been investigated. The apparent percentage of subunits engaged in complex formation was determined by incubation of radiolabeled cDNA probe with 50S subunits, followed by nitrocellulose membrane filtration of the reaction mixtures and measurement of the bound radiolabeled cDNA probes by liquid scintillation counting of the filters. The site(s) of hybridization were determined by digestion of the RNA in the RNA/DNA heteroduplex by RNase H. The results of this study indicated that single-stranded sequences, 2058-2062, 2448-2454, 2467-2483, and 2497-2505 were available for hybridization to cDNA probes. Bases 2489-2496, which have been postulated to be base paired with 2455-2461 were also accessible for hybridization.
对大肠杆菌50S亚基中23S rRNA V结构域特定序列与互补寡脱氧核糖核苷酸（cDNA）的可及性进行了研究。通过将放射性标记的cDNA探针与50S亚基一起孵育，然后对反应混合物进行硝酸纤维素膜过滤，并通过对滤膜进行液体闪烁计数来测定结合的放射性标记cDNA探针，从而确定参与复合物形成的亚基的表观百分比。通过用RNase H消化RNA/DNA异源双链体中的RNA来确定杂交位点。这项研究的结果表明，单链序列2058 - 2062、2448 - 2454、2467 - 2483和2497 - 2505可用于与cDNA探针杂交。据推测与2455 - 2461碱基配对的2489 - 2496碱基也可用于杂交。
