用于在金黄色葡萄球菌和肉葡萄球菌中克隆的质粒载体的构建与表征

Construction and characterization of plasmid vectors for cloning in Staphylococcus aureus and Staphylococcus carnosus.

作者信息

Keller G, Schleifer K H, Götz F

出版信息

Plasmid. 1983 Nov;10(3):270-8. doi: 10.1016/0147-619x(83)90041-0.

DOI:10.1016/0147-619x(83)90041-0

Abstract

Several plasmid vectors for cloning in Staphylococcus aureus and S. carnosus have been constructed and characterized. The chimeric plasmids are composed of parts of the following parental plasmids: The chloramphenicol-resistance plasmid, pC194, the tetracycline-resistance plasmid, pMK148, and the erythromycin-resistance plasmid, pE12. All the chimeric plasmids confer two selectable antibiotic-resistance markers on host cells. Insertional inactivation of the various antibiotic-resistance markers occurred at the BclI site of pE12, and the Sau96- or AvaII-site of pMK148; only a slight inactivation of the chloramphenicol-resistance marker occurred at the HaeIII-site of pC194. The chimeric plasmids pCT20 and pCE10 are both stable in S. aureus and S. carnosus. In addition, the hybrid plasmids of pCT20 and pCE10, containing lambda-DNA fragments in various restriction sites between 0.4 and 1.2 kb, are stably maintained. The inserted lambda-DNA fragments appear unchanged.

摘要

已构建并鉴定了几种用于在金黄色葡萄球菌和肉葡萄球菌中克隆的质粒载体。嵌合质粒由以下亲本质粒的部分组成：氯霉素抗性质粒pC194、四环素抗性质粒pMK148和红霉素抗性质粒pE12。所有嵌合质粒都赋予宿主细胞两种可选择的抗生素抗性标记。各种抗生素抗性标记的插入失活发生在pE12的BclI位点、pMK148的Sau96或AvaII位点；在pC194的HaeIII位点，氯霉素抗性标记仅发生轻微失活。嵌合质粒pCT20和pCE10在金黄色葡萄球菌和肉葡萄球菌中均稳定。此外，pCT20和pCE10的杂交质粒，在0.4至1.2 kb之间的各种限制性位点含有λ-DNA片段，能稳定维持。插入的λ-DNA片段似乎未发生变化。