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Excision de la region du DNA liee a la RNA polymerase in vitro
Author(s) -
Sentenac A.,
Ruet A.,
Fromageot P.
Publication year - 1968
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(68)80099-7
Subject(s) - microbiology and biotechnology , oligonucleotide , polymerase , nucleotide , dna , biology , rna , chemistry , dna polymerase , rna polymerase , deoxyribonuclease , biochemistry , gene
After synthesis of short, nascent oligonucleotide in the presence of ( 32 P)DNA, GTP, CTP, UTP and 3′dATP, one can excise with deoxyribonuclease a ternary complex of RNA polymerase, protected DNA and oligonucleotide, while the enzyme simply bound to the template is removed by increasing the ionic strength. This ternary complex is retained on nitrocellulose membranes. On polyacrylamide gel electrophoresis it migrates faster than RNA polymerase alone. The protected portion of the DNA is constituted of about 75 nucleotides. It might represent the sites for RNA initiation.