Gil J A, Kieser H M, Hopwood D A
Gene. 1985;38(1-3):1-8. doi: 10.1016/0378-1119(85)90197-0.
A gene (cat) for chloramphenicol (Cm) acetyltransferase (CAT) was cloned from Streptomyces acrimycini into S. lividans 66 on the plasmid vector pIJ61. The cat gene was localized on a 1.7-kb BclI fragment, which probably also carries the cat promoter. This DNA fragment conferred Cm resistance, through CAT activity, on S. lividans, S. coelicolor and S. parvulus, but not on Escherichia coli when inserted in the BamHI site of the tetracycline-resistance(TcR) gene of pBR322. However, when inserted in a particular orientation in this site, spontaneous deletions of 0.7 kb led to CAT activity and Cm resistance. DNA homologous to the 1.7-kb BclI cat fragment was found in most, but not all, of a series of other streptomycetes that have CAT activity. The cat provides a potentially useful screening marker for Streptomyces cloning vectors.
从尖锐链霉菌中克隆出一个编码氯霉素乙酰转移酶(CAT)的基因(cat),并通过质粒载体pIJ61将其导入变铅青链霉菌66中。cat基因定位在一个1.7kb的BclI片段上,该片段可能也携带cat启动子。当将此DNA片段插入pBR322的四环素抗性(TcR)基因的BamHI位点时,它通过CAT活性赋予变铅青链霉菌、天蓝色链霉菌和小链霉菌氯霉素抗性,但对大肠杆菌无此作用。然而,当以特定方向插入该位点时,0.7kb的自发缺失导致CAT活性和氯霉素抗性。在一系列具有CAT活性的其他链霉菌中,多数(但不是全部)发现了与1.7kb BclI cat片段同源的DNA。cat为链霉菌克隆载体提供了一个潜在有用的筛选标记。