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.
Sode
K
K
Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, Japan.
Hatano
N
N
Tatara
M
M
D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics*; D015342:DNA Probes; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005822:Genetic Vectors; D016680:Genome, Bacterial; D011401:Promoter Regions, Genetic; D013050:Spectrometry, Fluorescence
D015342:DNA Probes; D015500:Chloramphenicol O-Acetyltransferase