Abstract
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 flowthrough 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.
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Sode, K., Hatano, N. & Tatara, M. Cloning of a marine cyanobacterial promoter for foreign gene expression using a promoter probe vector. Appl Biochem Biotechnol 59, 349–360 (1996). https://doi.org/10.1007/BF02783576
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DOI: https://doi.org/10.1007/BF02783576