Expression of the Escherichia coli lacZ gene on a plasmid vector in a cyanobacterium

• Published in: Science (American Association for the Advancement of Science) 1985-11, Vol.230 (4727), p.805-807
• Main Authors: Buzby, J.S, Porter, R.D, Stevens, S.E. Jr
• Format: Article
• Language: English
• Subjects: AGMENELLUM QUADRUPLICATUM; Bacterial genetics; beta-Galactosidase - metabolism; Biological and medical sciences; Biotechnology; Cyanobacteria - genetics; CYANOPHYTA; DNA Restriction Enzymes; Enzymes; ESCHERICHIA; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; GENE; Gene expression; GENES; Genetic engineering; Genetic mutation; Genetic technics; Genetic variation; Genetic Vectors; Lac Operon; Liquids; Methods. Procedures. Technologies; MUTANT; MUTANTES; MUTANTS; Plasmid vectors; PLASMIDE; PLASMIDIOS; PLASMIDS; Streptococcus pneumoniae; transformation; Vectors (cloning, transfer, expression). Insertion sequences and transposons
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.2997920

A biphasic plasmid vector was used to introduce the Escherichia coli K-12 lac operon into the unicellular cyanobacterium Agmenellum quadruplicatum PR-6. The PR-6 transformants expressed bβ -galactosidase at nearly as high a level as did Escherichia coli transformants. In order to accomplish this, it was necessary to obtain PR-6 mutants that could be transformed by plasmids with unmodified recognition sites for the endogenous PR-6 restriction endonuclease Aqu I. These mutants were generated by a variation of the ectopic mutagenesis techniques that have been used in other naturally transforming bacteria. The ability to assay the expression of lacZ in PR-6 paves the way for the construction of gene fusions with various PR-6 promoters and quantitation of their expression under specific in vivo conditions.