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Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors

✍ Scribed by Miroslawa M. Bagdasarian; Egon Amann; Rudolf Lurz; Beate Rückert; Michael Bagdasarian

Book ID
118174970
Publisher
Elsevier Science
Year
1983
Tongue
English
Weight
964 KB
Volume
26
Category
Article
ISSN
0378-1119

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✦ Synopsis

A broad-host-range vector, pKT240, containing the structural gene (aph) for aminoglycoside phosphotransferase (APH), without promoter, has been constructed. Insertion of DNA fragments carrying promoters upstreamof the aph gene into the unique EcoRI site of this vector results in the expression of the aph gene and consequently the resistance of the host cells to streptomycin.

The new vector has been used to show that the hybrid trp-Zac (tat) promoter and the promoter of the la& gene of Escherichia coli are active in Pseudomonas putida.

Derivatives of pKT240 containing tat and Ia@ sequences may be used as wide-host-range expression vectors. Regulated overproduction of APH and catechol2,3-oxygenase can be obtained with the aid of the new vectors in both E. coli and P. putida.

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✍ John Kidwell; Dana Kolibachuk; Douglas Dennis 📂 Article 📅 1996 🏛 John Wiley and Sons 🌐 English ⚖ 738 KB

To determine the utility of coupling runaway replication to the expression of cloned genes under the control of strong promoters, lacZ transcriptional fusions to the trp or tac promoter (Ptrp or Ptac) were constructed using plasmids in which the copy number is thermally regulated. Cells containing t