Komiya H, Kawakami M, Takemura S
J Biochem. 1981 Mar;89(3):717-22. doi: 10.1093/oxfordjournals.jbchem.a133251.
A low molecular weight RNA fraction was obtained from the posterior silk glands of Bombyx mori. 5S rRNA was purified from this fraction by gel filtration on a column of Sephadex G-100 and electrophoresis on a discontinuous polyacrylamide gel devised for large scale preparation. The nucleotide sequence of this RNA was determined mainly by a chemical sequencing method on polyacrylamide gels using RNA labeled with [32P]pCp at the 3' terminus (1). An enzymatic sequencing method on gels using 32P-labeled RNA at the 5' end (2) and conventional sequence analyses of the complete digests of the unlabeled RNA with RNase T1 [EC 3.1.27.3] and RNase A [EC 3.1.27.5] were used for complete determination of the total sequence. 16 base substitutions were observed between B. mori and Drosophila melanogaster 5S rRNAs.
从家蚕的后部丝腺中获得了一个低分子量RNA组分。通过在Sephadex G - 100柱上进行凝胶过滤以及在为大规模制备设计的不连续聚丙烯酰胺凝胶上进行电泳,从该组分中纯化出了5S rRNA。该RNA的核苷酸序列主要通过使用在3'末端用[32P]pCp标记的RNA在聚丙烯酰胺凝胶上的化学测序方法来确定(1)。还使用了在凝胶上对5'末端用32P标记的RNA进行酶促测序方法(2),以及用RNase T1 [EC 3.1.27.3]和RNase A [EC 3.1.27.5]对未标记RNA的完全消化产物进行常规序列分析,以完整确定整个序列。在家蚕和黑腹果蝇的5S rRNA之间观察到16个碱基替换。