CLONING AND ANALYSIS OF THE PROMOTER REGION OF THE ERYTHROMYCIN-RESISTANCE GENE (ERME) OF STREPTOMYCES-ERYTHRAEUS

CLONING AND ANALYSIS OF THE PROMOTER REGION OF THE ERYTHROMYCIN-RESISTANCE GENE (ERME) OF STREPTOMYCES-ERYTHRAEUS
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DOI:
10.1016/0378-1119(86)90122-8
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发表时间:
1986-01-01
期刊:
影响因子:
3.5
通讯作者:
WARD, JM
WARD, JM
中科院分区:
生物学3区
文献类型:
--
作者:
BIBB, MJ;JANSSEN, GR;WARD, JM

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利用质粒载体pIJ 61将含有红霉素(Er)产生菌Streptomyceserythromycin(Er)抗性(ermE)基因的编码和调节序列的DNA片段克隆到链霉菌中。ermE在大肠杆菌中的亚克隆表达的研究推断了ermE的大致位置和方向。通过核苷酸序列测定、高分辨率S1和核酸外切酶VII图谱、体外转录和体内启动子探针分析,对链霉菌ermE基因的转录调控序列进行了研究。典型原核生物外观的串联启动子启动ermE编码区的转录;鉴定了类似序列的启动子,其启动可能的编码区的转录,该编码区与ermE的方向相反。有人建议,这些网站代表一类植物表达的链霉菌启动子,利用的RNA聚合酶全酶的形式,也认识到其他细菌属的典型启动子。
A DNA fragment containing the coding and regulatory sequences of the erythromycin (Er) resistance (ermE) gene of the Er producer Streptomyces erythraeus was cloned in Streptomyces 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 recognises typical promoters of other bacterial genera.