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Nucleic Acids Research, 1979, Vol. 7, No. 6 1621-1633
© 1979

Articles

RNA-linked nascent DNA pieces in phage T7-infected Escherichia coli. III. Detection of intact primer RNA

Tohm Ogawa and Tuneko Okazaki

Institute of Molecular Biology, Faculty of Science, Nagoya University Nagoya 464, Japan

Received August 10, 1979. RNA-linked DNA fragments of T7-lnfected Escherlchla coli were labeled with [32P]orthophosphate In vivo. The RNA segments of the labeled fragments were Isolated by degrading the DNA portion with the 3'-> 51 exo-nuclease intrinsic to bacterlophage T4 DNA polymerase and fractionated according to net charge by a DEAE-Sephadex A-25 column chromatography in the presence of 7 M urea. Tri-, tetra- and pentanucleotldes were obtained which have ATP residues at their 5' ends. Most of the pentanucleotides had a single deoxynucleotlde at the 3' end but a minor portion was totally an ollgoribonucleotlde. In the light of prior results, the former is a cooligomer of an intact tetraribonucleotide primer and a monodeoxynucleotide and the latter is an intact pentarlbonucleotide primer. Tri- and tetraribonucleotides with ATP at the 5' ends had no deoxynucleotide at the 3' ends, therefore it is not clear if intact triribonucleotide primers are present. The 5'-terminal di-nucleotides of the tetra- and pentanucleotides were mostly pppApC and a trace amount of pppApA was present.


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 Cold Spring Harb Symp Quant BiolHome page
C.W. Fuller, B.B. Beauchamp, M.J. Engler, R.L. Lechner, S.W. Matson, S. Tabor, J.H. White, and C.C. Richardson
Mechanisms for the Initiation of Bacteriophage T7 DNA Replication
Cold Spring Harb Symp Quant Biol, January 1, 1983; 47(0): 669 - 679.
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