Wilson C R, Baldwin N J
J Bacteriol. 1978 Oct;136(1):402-13. doi: 10.1128/jb.136.1.402-413.1978.
Four chloramphenicol resistance (Cm) and four tetracycline resistance (Tc) plasmids from Staphylococcus aureus were characterized by restriction endonuclease mapping. All four Tc plasmids had molecular masses of 2.9 megadaltons (Mdaltons) and indistinguishable responses to seven different restriction endonucleases. The four Cm plasmids (pCW6, pCW7, pCW8, and pC221) had molecular masses of 2.6, 2.8, 1.9, and 2.9 Mdaltons, respectively. The four Cm plasmids also differed both in the level of resistance to Cm and in susceptibility to retriction endonucleases. Single restriction endonuclease sites contained within each plasmid included the following: in pCW6 for HindIII, XbaI, HpaII, and BstEII; in pCW7 for HindIII, BstEII, BglII, HaeIII, and HpaII; in pCW8 for HindIII, HaeIII, and HpaII; in pC221 for HindIII, BstEII, and EcoRI. The molecular cloning capabilities of pCW8 and pC221 were determined. Cm and erythromycin resistance (Em) recombinant plasmids pCW12, PCW13, and pCW14 were constructed and used to transform S. aureus 8325-4. A 2.8-Mdalton HindIII fragment from plasmid pI258 was found to encode Em resistance and contain single sites for the retriction endonucleases BglII, PstI, HaeIII, and HpaII. The largest EcoRI fragment (8 Mdaltons) from pI258 contained the HindIII fragment encoding Em resistance intact. Cloning of DNA into the BglII site of pCW14 did not alter Em resistance. Cloning of DNA into the HindIII site of pCW8 and the HindIII and EcoRI sites of pC221 did not disrupt either plasmid replication of Cm resistance.
通过限制性内切酶图谱分析对来自金黄色葡萄球菌的4个氯霉素抗性（Cm）质粒和4个四环素抗性（Tc）质粒进行了表征。所有4个Tc质粒的分子量均为2.9兆道尔顿（Mdaltons），并且对7种不同的限制性内切酶有无法区分的反应。4个Cm质粒（pCW6、pCW7、pCW8和pC221）的分子量分别为2.6、2.8、1.9和2.9 Mdaltons。这4个Cm质粒在对Cm的抗性水平以及对限制性内切酶的敏感性方面也存在差异。每个质粒中包含的单限制性内切酶位点如下：pCW6中含有HindIII、XbaI、HpaII和BstEII的位点；pCW7中含有HindIII、BstEII、BglII、HaeIII和HpaII的位点；pCW8中含有HindIII、HaeIII和HpaII的位点；pC221中含有HindIII、BstEII和EcoRI的位点。测定了pCW8和pC221的分子克隆能力。构建了Cm和红霉素抗性（Em）重组质粒pCW12、PCW13和pCW14，并用于转化金黄色葡萄球菌8325 - 4。发现来自质粒pI258的一个2.8 - Mdaltons HindIII片段编码Em抗性，并且含有限制性内切酶BglII、PstI、HaeIII和HpaII的单一位点。来自pI258的最大EcoRI片段（8 Mdaltons）完整地包含了编码Em抗性的HindIII片段。将DNA克隆到pCW14的BglII位点不会改变Em抗性。将DNA克隆到pCW8的HindIII位点以及pC221的HindIII和EcoRI位点不会破坏质粒复制或Cm抗性。
