TY - JOUR
T1 - Separation of α- and β-globin messenger RNAs by formamide gel electrophoresis
AU - Kazazian, Haig H.
AU - Snyder, Pamela G.
AU - Tu-chen, Cheng
N1 - Funding Information:
This work was supported in part by NIH research grant AM 13983.
PY - 1974/8/5
Y1 - 1974/8/5
N2 - Electrophoresis on 7.5% polyacrylamide gels in 98% formamide splits the 10s RNA of EDTA-treated polysomes from rabbit reticulocytes into two bands of similar intensity. Fractions containing first 10s RNA1, both 10s RNAs, and nearly-pure second 10s RNA were obtained and tested in a cell-free system derived from ascites tumor cells. These RNA preparations directed the synthesis of product composed of 85-100% α chains, 55-70% β chains, and 75-87% β chains, respectively. The relative amounts of the two 10s RNAs in certain fractions were determined by formamide electrophoresis and correlated well with the relative synthesis of β and α chains directed by these fractions in the cell-free system. Therefore, the α and β globin mRNAs are separated by this procedure. Moreover, electrophoresis of globin mRNA further purified by oligo-dT cellulose chromatography indicates that the concentration of α mRNA exceeds that of β mRNA on rabbit reticulocytes polysomes.
AB - Electrophoresis on 7.5% polyacrylamide gels in 98% formamide splits the 10s RNA of EDTA-treated polysomes from rabbit reticulocytes into two bands of similar intensity. Fractions containing first 10s RNA1, both 10s RNAs, and nearly-pure second 10s RNA were obtained and tested in a cell-free system derived from ascites tumor cells. These RNA preparations directed the synthesis of product composed of 85-100% α chains, 55-70% β chains, and 75-87% β chains, respectively. The relative amounts of the two 10s RNAs in certain fractions were determined by formamide electrophoresis and correlated well with the relative synthesis of β and α chains directed by these fractions in the cell-free system. Therefore, the α and β globin mRNAs are separated by this procedure. Moreover, electrophoresis of globin mRNA further purified by oligo-dT cellulose chromatography indicates that the concentration of α mRNA exceeds that of β mRNA on rabbit reticulocytes polysomes.
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U2 - 10.1016/S0006-291X(74)80086-0
DO - 10.1016/S0006-291X(74)80086-0
M3 - Article
C2 - 4415727
AN - SCOPUS:0016184828
VL - 59
SP - 1053
EP - 1061
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -