Highly specific transcription of globin sequences in isolated reticulocyte nuclei

Chicken reticulocyte nuclei have been isolated under conditions where the endogenous RNA polymerase activity is retained and catalyzes the incorporation of ribonucleotide triphosphates into RNA linearly for at least 5 hours at 25°C after an initial burst of activity. Transcription has also been carr...

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Bibliographic Details
Published inBiochemical and biophysical research communications Vol. 77; no. 4; pp. 1478 - 1485
Main Authors Fodor, Eric J.B., Doty, Paul
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 22.08.1977
Subjects
Amanitins - pharmacology
Animals
Cell Nucleus - drug effects
Cell Nucleus - metabolism
Chickens
Globins - biosynthesis
Guanosine Triphosphate - metabolism
Kinetics
Male
Mercury - pharmacology
Nucleic Acid Hybridization
Reticulocytes - metabolism
RNA - biosynthesis
Transcription, Genetic - drug effects
cDNA
SDS
EDTA
HEPES
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Summary:Chicken reticulocyte nuclei have been isolated under conditions where the endogenous RNA polymerase activity is retained and catalyzes the incorporation of ribonucleotide triphosphates into RNA linearly for at least 5 hours at 25°C after an initial burst of activity. Transcription has also been carried out utilizing 5-mercurated uridine triphosphate. Hybridization of the mercurated RNA, isolated from the endogenous RNA by affinity chromatography on sulfhydryl Sepharose with globin cDNA reveals that 0.24 percent of the transcript is globin sequence: this is about 100-fold higher than that reported for chromatin with E. coli RNA polymerase.
ISSN:0006-291X
1090-2104
DOI:10.1016/S0006-291X(77)80145-9