Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography.

R. K. Patient, S. C. Hardies, J. E. Larson, R. B. Inman, L. E. Maquat, R. D. Wells

Research output: Contribution to journalArticle

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Abstract

The properties of the fractionated Hae III fragments of pRZ2 DNA (Patient, R.K., Hardies, S.C., and Wells, R.D. (1979) J. Biol. Chem. 254, 5542-5547) were studied in an effort to determine why several of the fragments bind more tightly to RPC-5 than expected on the basis of their length. The purified fragments were analyzed for their nucleotide composition by direct determination of their constituent mononucleotides and by analytical CsCl and Cs2SO4 density gradient analyses. A-T-rich fragments elute at higher salt concentrations than fragments of equivalent size which are not A-T-rich. In addition, denaturation mapping studies by electron microscopy indicate that an A-T-rich run within an otherwise G-C-rich fragment can give rise to delayed elution. At least one other factor influences the separation of DNA restriction fragments by RPC-5 chromatography. Some of the fragments in this digest which elute later than predicted from their size either contain known genetic regulatory sites or bind regulatory proteins.

Original languageEnglish (US)
Pages (from-to)5548-5554
Number of pages7
JournalJournal of Biological Chemistry
Volume254
Issue number12
StatePublished - Jun 25 1979
Externally publishedYes

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Column chromatography
Fractionation
Chromatography
Denaturation
DNA
Electron microscopy
Electron Microscopy
Nucleotides
Salts
Chemical analysis
Proteins
cesium chloride

ASJC Scopus subject areas

  • Biochemistry

Cite this

Patient, R. K., Hardies, S. C., Larson, J. E., Inman, R. B., Maquat, L. E., & Wells, R. D. (1979). Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography. Journal of Biological Chemistry, 254(12), 5548-5554.

Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography. / Patient, R. K.; Hardies, S. C.; Larson, J. E.; Inman, R. B.; Maquat, L. E.; Wells, R. D.

In: Journal of Biological Chemistry, Vol. 254, No. 12, 25.06.1979, p. 5548-5554.

Research output: Contribution to journalArticle

Patient, RK, Hardies, SC, Larson, JE, Inman, RB, Maquat, LE & Wells, RD 1979, 'Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography.', Journal of Biological Chemistry, vol. 254, no. 12, pp. 5548-5554.
Patient RK, Hardies SC, Larson JE, Inman RB, Maquat LE, Wells RD. Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography. Journal of Biological Chemistry. 1979 Jun 25;254(12):5548-5554.
Patient, R. K. ; Hardies, S. C. ; Larson, J. E. ; Inman, R. B. ; Maquat, L. E. ; Wells, R. D. / Influence of A-T content on the fractionation of DNA restriction fragments by RPC-5 column chromatography. In: Journal of Biological Chemistry. 1979 ; Vol. 254, No. 12. pp. 5548-5554.
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