PMID: 9443386Jan 27, 1998Paper

Purification and properties of a DNA primase from Nicotiana tabacum

Planta
M M Garcia-Maya, K W Buck

Abstract

A DNA primase was isolated from a nuclear fraction from leaves of tobacco (Nicotiana tabacum L. cv. Samsun) and from purified nuclei prepared from tobacco suspension culture cells. The DNA primase was purified to homogeneity (i) for preparations from leaves, by ammonium sulphate fractionation, followed by chromatography on columns of phosphocellulose, Q-Sepharose, heparin-Sepharose and single-stranded DNA cellulose, and sedimentation in a glycerol gradient, or (ii) for preparations from cells, by chromatography on single-stranded DNA cellulose, followed by ammonium sulphate precipitation and chromatography on columns of High Q, heparin-Sepharose and Mono Q. In glycerol gradients, the DNA primase sedimented at a rate corresponding to a molecular mass of about 120 kDa. In SDS-polyacrylamide gel electrophoresis, the primase was resolved into two polypeptide subunits of 63 kDa and 53 kDa, which are similar in size to the primase subunits of animal and yeast DNA polymerase alpha-primase complexes. On poly(dT) or phage M13 single-stranded DNA templates, the DNA primase catalysed the synthesis of oligoribonucleotides up to 20 nucleotides in length, which could serve as primers for DNA synthesis catalysed by Escherichia coli DNA polyme...Continue Reading

Citations

May 23, 2008·Critical Reviews in Plant Sciences·Miguel Garcia-Diaz, Katarzyna Bebenek
Apr 3, 2001·Journal of Experimental Botany·J A BryantS J Aves
Feb 14, 2008·The Plant Journal : for Cell and Molecular Biology·Jan A Miernyk, Jay J Thelen
Dec 21, 2010·Annals of Botany·Wojciech Strzalka, Alicja Ziemienowicz
Aug 27, 1999·Archives of Biochemistry and Biophysics·Z Nie, M Wu

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