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Applied Biochemistry and Biotechnology, Vol.59, No.3, 349-360, 1996
Cloning of a Marine Cyanobacterial Promoter for Foreign Gene-Expression Using a Promoter Probe Vector
A marine cyanobacterial promoter was cloned to allow efficient foreign gene expression. This was carried out using chloramphenicol acetyl transferase (CAT) as a marker protein. For rapid and simple measurement of CAT activity, a method based on a fluorescently labeled substrate was improved by utilizing HPLC equipped with a flow-through fluorescent spectrophotometer. This method was used in conjunction with a newly constructed promoter probe vector. Cyanobacterial transformants, harboring plasmid containing a cloned 2-kbp marine cyanobacterial genomic fragment, showed a 10-fold higher CAT activity, compared with that achieved using the kanamycin-resistant gene promoter. From the sequence analysis of the cloned fragment, a putative promoter region was found.
Keywords:BROAD HOST RANGE;CHLORAMPHENICOL ACETYLTRANSFERASE;UNICELLULAR CYANOBACTERIA;COPY NUMBER;SYSTEM