Cloning of a marine cyanobacterial promoter for foreign gene expression using a promoter probe vector

Journal Article · Sat Jun 01 00:00:00 EDT 1996 · Applied Biochemistry and Biotechnology

DOI:https://doi.org/10.1007/BF02783576· OSTI ID:588692

Sode, Koji; Hatano, Naoaki; Tatara, Masahiro ^[1]

---

1. Tokyo Univ. of Agriculture and Technology (Japan)

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. 20 refs., 7 figs., 2 tabs.

• Sponsoring Organization: USDOE
• OSTI ID: 588692
• Journal Information: Applied Biochemistry and Biotechnology, Vol. 59, Issue 3; Other Information: PBD: Jun 1996
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
08 HYDROGEN FUEL
54 ENVIRONMENTAL SCIENCES
CHLORAMPHENICOL
DNA-CLONING
GENES
GENE REGULATION
CYANOBACTERIA
GENE REPRESSORS
GENETIC ENGINEERING
PRODUCTION
HYDROGEN
NUCLEOTIDES
BIOLOGICAL MARKERS
PROTEINS
SPECTROPHOTOMETRY
FLUORESCENCE
PLASMIDS
DNA SEQUENCING
SOLAR ENERGY CONVERSION
CARBON DIOXIDE
CARBON DIOXIDE FIXATION