The Separation and Identification of α- and β-Globin Messenger Ribonucleic Acids

作者：Marcelle R. Morrison, Shirley A. Brinkley, Jack Gorski, Jerry B. Lingrel · 发表于：Journal of Biological Chemistry · 年份：1974 · DOI：10.1016/s0021-9258(19)42362-4 · 被引用次数：60 · 研究领域：DNA and Nucleic Acid Chemistry

The size heterogeneity of the mouse and rabbit globin messenger ribonucleic acids (mRNAs) purified by oligodeoxythymidylate-cellulose has been studied by polyacrylamide gel electrophoresis. The globin mRNAs migrate as a diffuse band on sodium dodecyl sulfate 6% polyacrylamide gels with no separation of the α and β messengers. The diffuse migration pattern is partly due to secondary structure effects as electrophoresis on 4% polyacrylamide gels in the presence of formamide partially resolves two bands in the 9 S RNA region. Increasing the concentration of polyacrylamide to 10% further resolves these two bands. The difference in molecular weight between the bands is greater than that expected on the basis of a difference in the coding sequence of 15 nucleotides between the α- and β-globin mRNAs. To identify the bands, α-globin mRNA was isolated from the rabbit reticulocyte post-ribosomal supernatant fraction and subjected to electrophoresis along with total rabbit globin mRNA on the 10% polyacrylamide-formamide gels. The α-globin mRNA migrated with the faster moving band of the total mRNA preparation, thus identifying this band as the α-globin mRNA and, by inference, the slower migrating band as the β-globin mRNA. Using mRNA preparations enriched in one or the other of the messengers, it was shown that the mouse α- and β-globin mRNAs migrate identically with those of rabbit. The identity of the two bands in mouse mRNAs was further established by determining their ability to dir...