Bibb M J, Janssen G R, Ward J M
Gene. 1985;38(1-3):215-26. doi: 10.1016/0378-1119(85)90220-3.
A DNA fragment containing the coding and regulatory sequences of the erythromycin (Er) resistance (ermE) gene of the Er produces Streptomyces erythraeus was cloned in Streptomyces lividans using the plasmid vector pIJ61. The approximate location and orientation of ermE were deduced from studies of its expression after subcloning in Escherichia coli. Sequences responsible for transcription of ermE in Streptomyces were studied by nucleotide (nt) sequencing, high resolution S1 and exonuclease VII mapping, in vitro transcription and in vivo promoter-probing. Tandemly arranged promoters of typical prokaryotic appearance initiate transcription of the coding region of ermE; a promoter of similar sequence was identified that initiates transcription of a likely coding region running in the opposite direction to ermE. It is suggested that these sites represent a class of vegetatively expressed Streptomyces promoter that is utilised by a form of RNA polymerase holoenzyme that also recognizes typical promoters of other bacterial genera.
使用质粒载体pIJ61，将含有产生红霉素的红色糖多孢菌红霉素（Er）抗性（ermE）基因编码和调控序列的DNA片段克隆到淡青链霉菌中。通过在大肠杆菌中进行亚克隆后对其表达的研究，推断出ermE的大致位置和方向。通过核苷酸（nt）测序、高分辨率S1和核酸外切酶VII图谱分析、体外转录和体内启动子探测，研究了链霉菌中负责ermE转录的序列。具有典型原核生物外观的串联排列启动子启动ermE编码区的转录；鉴定出一个序列相似的启动子，它启动了一个可能与ermE方向相反的编码区的转录。有人认为，这些位点代表了一类营养表达的链霉菌启动子，它们被一种RNA聚合酶全酶形式所利用，这种全酶也能识别其他细菌属的典型启动子。
