Bardonnet N, Trautwetter A, Couchoux-Luthaud G, Blanco C
Laboratoire de Microbiologie (CNRS UM 380024), I.N.S.A., Villeurbanne, France.
Mol Gen Genet. 1988 May;212(2):390-2. doi: 10.1007/BF00334714.
We have constructed promoter-probe plasmids, pNB4 and pNB5, based on the promoterless gene for beta-glucuronidase (uidA) of Escherichia coli. Unique restriction sites for EcoRI, SacI, KpnI, SmaI, XmaI, XbaI, SalI, SphI and HindIII in pNB4 and for HindIII, PstI and BglII in pNB5 were included upstream of the uidA structural gene. The usefulness of these plasmids was demonstrated by cloning the promoter-operator region of the E. coli uxaB gene. We observed that expression of the uxaB-uidA operon fusion followed the transcription-regulating properties of the uxaB promoter. Another construct, pNB2, can be used to detect operator and terminator signals. Recipient cells transformed with such recombinant plasmids can be revealed by growth on medium containing a chromogenic beta-glucuronidase substrate.
我们基于大肠杆菌β-葡萄糖醛酸酶（uidA）的无启动子基因构建了启动子探针质粒pNB4和pNB5。在pNB4中，EcoRI、SacI、KpnI、SmaI、XmaI、XbaI、SalI、SphI和HindIII的独特限制性酶切位点以及在pNB5中HindIII、PstI和BglII的独特限制性酶切位点被包含在uidA结构基因的上游。通过克隆大肠杆菌uxaB基因的启动子-操纵子区域，证明了这些质粒的实用性。我们观察到uxaB-uidA操纵子融合体的表达遵循uxaB启动子的转录调控特性。另一个构建体pNB2可用于检测操纵子和终止子信号。用此类重组质粒转化的受体细胞可通过在含有显色性β-葡萄糖醛酸酶底物的培养基上生长来揭示。
