Stein D C, Silver L E, Clark V L, Young F E
J Bacteriol. 1984 May;158(2):696-700. doi: 10.1128/jb.158.2.696-700.1984.
DNA from Neisseria gonorrhoeae KH45 was partially digested with Sau3A and inserted into the BamHI site of the cloning vector pLES2 . After introduction into Escherichia coli JM83 by transformation, two different size classes of plasmids were isolated that could complement the proAB deletion of JM83 . These plasmids ( pLES4 and pLES7 ) were characterized by restriction endonuclease digestion. Southern hybridization demonstrated that the inserts had sequence homology. Various deletions of these plasmids were constructed that had lost the ability to complement the proA lesion of chi 463, the proB lesion of chi 340, or both (plasmids pLES9 , pLES8 , and pLES10 , respectively). These deleted plasmids were introduced into a proline-requiring strain of N. gonorrhoeae, F62, with plasmids pLES4 , pLES7 , and pLES8 possessing the ability to correct the proline requirement of F62. Further analysis indicated that the hybrid plasmids were stably maintained as plasmids in N. gonorrhoeae.
用Sau3A对淋病奈瑟菌KH45的DNA进行部分酶切，然后插入到克隆载体pLES2的BamHI位点。通过转化将其导入大肠杆菌JM83后，分离出两种不同大小的质粒，它们能够互补JM83的proAB缺失。这些质粒（pLES4和pLES7）通过限制性内切酶消化进行表征。Southern杂交表明插入片段具有序列同源性。构建了这些质粒的各种缺失突变体，它们分别丧失了互补chi 463的proA缺陷、chi 340的proB缺陷或两者缺陷的能力（分别为质粒pLES9、pLES8和pLES10）。将这些缺失质粒导入需要脯氨酸的淋病奈瑟菌菌株F62中，其中质粒pLES4、pLES7和pLES8能够校正F62对脯氨酸的需求。进一步分析表明，这些杂交质粒在淋病奈瑟菌中作为质粒稳定维持。
