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Nucleic Acids Research, 1974, Vol. 1, No. 11 1521-1530
© 1974

Articles

Nucleotide sequences of the 5' termini of øX174 mRNAs synthesized in vitro

Lloyd H. Smith, Karel Grohmann and R.L. Sinsheimer

Division of Biology, California Institute of Technology Pasadena, California 91109, USA

Received August 20, 1974. When using øX17U RFI DNA as a template, in vitro, E. coli RNA polymerase synthesizes four major purine triphosphate-containing 5' end sequences. RNase A digests of {alpha}32P labeled RNA were further digested with spleen exonuclease to remove the bulk of the oligonucleotides with 5' hydroxyls and then chromatographed on DEAE cellulose to resolve the remaining 5' terminal oligonucleotides. By application of standard separation and sequence techniques, the major 5' end sequences were shown to be: pppApUp(Cp), pppApApApUp(Cp), pppApApApApUp(Cp), and pppGpApUp(Gp).


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