DNA synthesis catalyzed in vitro by yeast extracts using A 2 μm DNA containing plasmid as template for enzymatic DNA synthesis

Summary Partially purified cell-free extracts prepared from growing cells of the yeast Saccharomyces cerevisiae catalyze DNA synthesis directed by the chimeric plasmid BTYP-2 containing the whole 2 m DNA sequence. The biochemical properties of the reaction have been examined and inactivation studies indicate that DNA synthesis is only due to protein factors. Analysis of the products of the DNA synthesis reaction demonstrates that part of the in vitro synthesized DNA is not covalently linked to the template, thus suggesting initiation of new DNA chains. The analysis of the distribution of the labelled products at early times of reaction showed a preferential synthesis on the 2 m portion of the BTYP-2 DNA template.Other abbreviations used are BSA bovine serum albumin - DTT dithiotreitol - EDTA ethylendiaminetetraacetate - EGTA ethylenglycol-bis(-aminoethyl-ether)N,N-tetraacetic acid PhcMeSO₂F, phenylmethylsulfonylfluoride - SDS sodium dodecyl sulfateAbbreviations: The abbreviations used are those recommended by the IUPAC-IUB Commission on Biochemical Nomenclature (CBN).