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. 1979 Nov 24;7(6):1621–1633. doi: 10.1093/nar/7.6.1621

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

Tohru Ogawa 1, Tuneko Okazaki 1
PMCID: PMC342332  PMID: 388359

Abstract

RNA-linked DNA fragments of T7-infected Escherichiacoli were labeled with [32P]orthophosphate invivo. The RNA segments of the labeled fragments were isolated by degrading the DNA portion with the 3′→ 5′ exonuclease intrinsic to bacteriophage 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 pentanucleotides were obtained which have ATP residues at their 5′ ends. Most of the pentanucleotides had a single deoxynucleotide at the 3′ end but a minor portion was totally an oligoribonucleotide. 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 pentaribonucleotide 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 dinucleotides of the tetra- and pentanucleotides were mostly pppApC and a trace amount of pppApA was present.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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