Manch J N, Crawford I P
J Bacteriol. 1981 Apr;146(1):102-7. doi: 10.1128/jb.146.1.102-107.1981.
The Pseudomonas aeruginosa tryptophan synthase genes, trpA and trpB, which are induced by their substrate indoleglycerol phosphate, were cloned along with their controlling region into the BamHI site of pBR322 to produce the 10.7-megadalton plasmid pZAZ5. SalI partial digestion and ligation yielded a smaller plasmid, pZAZ167, with the chromosomal insert reduced in size from 8.1 to 3.4 megadaltons. Both pZAZ5 and pZAZ167 display Pseudomonas-like regulation of the trpA and trpB genes. Deletion of an EcoRI fragment or a BglII fragment from pZAZ167 yielded plasmids pZAZ168 and pZAZ169; the former expresses trpB but not trpA, and the latter has lost both activities. A deleted form of pZAZ5 designated pZAZ101 was obtained by excising a BglII-BamHI segment and religating the trip gene segment in the opposite orientation. This plasmid expresses trpA and trpB constitutively. The physical maps of these plasmids establish the gene order: promoter-trpB-trpA.
铜绿假单胞菌色氨酸合成酶基因trpA和trpB受其底物吲哚甘油磷酸诱导，它们与其控制区一起被克隆到pBR322的BamHI位点，产生了10.7兆道尔顿的质粒pZAZ5。用SalI进行部分酶切和连接得到了一个较小的质粒pZAZ167，其染色体插入片段的大小从8.1兆道尔顿减少到3.4兆道尔顿。pZAZ5和pZAZ167都表现出对trpA和trpB基因的假单胞菌样调控。从pZAZ167中缺失一个EcoRI片段或一个BglII片段，得到质粒pZAZ168和pZAZ169；前者表达trpB但不表达trpA，后者则失去了这两种活性。通过切除一个BglII - BamHI片段并以相反方向重新连接trp基因片段，获得了一种缺失形式的pZAZ5，命名为pZAZ101。该质粒组成型表达trpA和trpB。这些质粒的物理图谱确定了基因顺序：启动子 - trpB - trpA。
