Kop J, Kopylov A M, Magrum L, Siegel R, Gupta R, Woese C R, Noller H F
J Biol Chem. 1984 Dec 25;259(24):15287-93.
A majority (approximately 89%) of the nucleotide sequence of Bacillus brevis 16 S rRNA has been determined by a combination of RNA sequencing methods. Several experimental approaches have been used to probe its structure, including (a) partial RNase digestion of 30 S ribosomal subunits, followed by two-dimensional native/denatured gel electrophoresis, in which base-paired fragments were directly identified; (b) identification of positions susceptible to cleavage by RNase A and RNase T1 in 30 S subunits; (c) sites of attack by cobra venom RNase on naked 16 S rRNA; and (d) nucleotides susceptible to attack by bisulfite in 16 S rRNA. These data are discussed with respect to a secondary structure model for B. brevis 16 S rRNA derived by comparative sequence analysis.
通过多种RNA测序方法相结合,已确定了短短芽孢杆菌16S rRNA约89%的核苷酸序列。已采用多种实验方法来探测其结构,包括:(a) 对30S核糖体亚基进行部分核糖核酸酶消化,然后进行二维天然/变性凝胶电泳,其中碱基配对片段可直接鉴定;(b) 鉴定30S亚基中易被核糖核酸酶A和核糖核酸酶T1切割的位置;(c) 眼镜蛇毒核糖核酸酶对裸露的16S rRNA的攻击位点;以及(d) 16S rRNA中易被亚硫酸氢盐攻击的核苷酸。结合通过比较序列分析得出的短短芽孢杆菌16S rRNA二级结构模型对这些数据进行了讨论。