Strzelecki A T, Goodman A E, Rogers P L
Department of Biotechnology, University of New South Wales, Australia.
Plasmid. 1987 Jul;18(1):46-53. doi: 10.1016/0147-619x(87)90077-1.
The stability of the broad-host-range IncW R plasmid Sa in Zymomonas mobilis ZM6100(Sa) was monitored using three antibiotic resistance markers carried by Sa. When grown in batch culture without selection, ZM6100(Sa) rapidly lost the Sa plasmid. When grown with selection for either kanamycin or spectinomycin resistance, the three Sa markers were retained in at least 90% of the population, with spontaneous loss of chloramphenicol resistance being observed in the rest of the population. When ZM6100(Sa) was grown with selection for chloramphenicol resistance, the Sa plasmid either formed a cointegrate plasmid, designated pNSW301, with a native ZM6100 plasmid or was lost except for the chloramphenicol resistance marker. Possible transportation of the chloramphenicol resistance region of Sa to the Z. mobilis chromosome was indicated by hybridization data. The cointegrate plasmid, pNSW301, was maintained stably, except for spontaneous loss of chloramphenicol resistance, in Z. mobilis without selection for 300 generations. pNSW301 was able to replicate in Escherichia coli and was capable of conjugal transfer.
利用由质粒Sa携带的三种抗生素抗性标记,监测了广宿主范围的IncW R质粒Sa在运动发酵单胞菌ZM6100(Sa)中的稳定性。在无选择压力的分批培养中生长时,ZM6100(Sa)迅速丢失质粒Sa。在有卡那霉素或壮观霉素抗性选择压力下生长时,三种Sa标记在至少90%的群体中得以保留,其余群体中观察到氯霉素抗性的自发丢失。当ZM6100(Sa)在有氯霉素抗性选择压力下生长时,质粒Sa要么与运动发酵单胞菌ZM6100的天然质粒形成一个共整合质粒,命名为pNSW301,要么除氯霉素抗性标记外全部丢失。杂交数据表明,Sa的氯霉素抗性区域可能转移到了运动发酵单胞菌染色体上。共整合质粒pNSW301在无选择压力的运动发酵单胞菌中稳定维持了300代,不过会自发丢失氯霉素抗性。pNSW301能够在大肠杆菌中复制,并且能够进行接合转移。