Cloning of a marine cyanobacterial promoter for foreign gene expression using a promoter probe vector |
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Authors: | Koji Sode Naoaki Hatano Masahiro Tatara |
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Institution: | (1) Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei-shi, 184 Tokyo, Japan |
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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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Keywords: | Marine cyanobacteria promoter foreign gene expression fluorescent CAT assay |
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