Department of Food Microbiology and National Food Biotechnology Centre, University College, Cork, Ireland.
Appl Environ Microbiol. 1990 Jan;56(1):195-201. doi: 10.1128/aem.56.1.195-201.1990.
A combination of plasmid curing and DNA-DNA hybridization data facilitated the identification of proteinase plasmids of 75 (pCI301) and 35 kilobases (pCI203) in the multi-plasmid-containing strains Lactococcus lactis subsp. lactis UC317 and L. lactis subsp. cremoris UC205, respectively. Both plasmids were transferred by conjugation to a plasmid-free background only after introduction of the conjugative streptococcal plasmid, pAMbeta1. All Prt transconjugants from matings involving either donor contained enlarged recombinant Prt plasmids. UC317-derived transconjugants were separable into different classes based on the presence of differently sized cointegrate plasmids and on segregation of the pCI301-derived Lac and Prt markers. All UC205-derived transconjugants harbored a single enlarged plasmid that was a cointegrate between pCI203 and pAMbeta1. The identification of prt genes on pCI301 and pCI203 derivatives was achieved by a combination of restriction enzyme and hybridization analyses.
质粒消除和 DNA-DNA 杂交数据的结合有助于鉴定含有多种质粒的乳球菌乳亚种 UC317 和乳球菌 cremoris UC205 中的蛋白酶质粒,分别为 75(pCI301)和 35 千碱基(pCI203)。只有在引入可接合的链球菌质粒 pAMbeta1 后,这两种质粒才能通过接合转移到无质粒背景。涉及任一供体的 Prt 转导子的所有交配都包含扩大的重组 Prt 质粒。UC317 衍生的转导子可以根据不同大小的共整合质粒的存在以及 pCI301 衍生的 Lac 和 Prt 标记的分离,分为不同的类别。所有 UC205 衍生的转导子都含有一个单一的扩大质粒,它是 pCI203 和 pAMbeta1 之间的共整合体。通过限制酶和杂交分析的组合,鉴定了 pCI301 和 pCI203 衍生物上的 prt 基因。