Pérez-González J A, Jiménez A
Biochem Biophys Res Commun. 1984 Dec 28;125(3):895-901. doi: 10.1016/0006-291x(84)91367-6.
The paromomycin producing organism Streptomyces rimosus forma paromomycinus is resistant to this antibiotic and contains a phosphotransferase which inactivates paromomycin. The gene encoding this enzyme has been inserted in the Streptomyces vector pIJ702 and then cloned in Streptomyces lividans, selecting for paromomycin-resistance. Three plasmids have been isolated and one of them, pMJ1, contains a 2.2 kb insert with a single HindIII restriction site. Insertion of foreign DNA in this site blocks the expression of the phosphotransferase enzyme indicating that it is within the cloned gene. These findings provide a new dominant selective marker for Streptomyces cloning vectors with the versatility of insertional inactivation.
产生巴龙霉素的微生物龟裂链霉菌巴龙霉素变种对这种抗生素具有抗性，并含有一种使巴龙霉素失活的磷酸转移酶。编码该酶的基因已被插入链霉菌载体pIJ702中，然后克隆到变铅青链霉菌中，选择对巴龙霉素具有抗性的菌株。已分离出三种质粒，其中一种pMJ1含有一个2.2 kb的插入片段，带有一个单一的HindIII限制性酶切位点。将外源DNA插入该位点会阻断磷酸转移酶的表达，这表明它位于克隆基因内。这些发现为链霉菌克隆载体提供了一种新的显性选择标记，具有插入失活的通用性。
