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

Appl Biochem Biotechnol. 1996 Jun;59(3):349-60. doi: 10.1007/BF02783576.

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chloramphenicol O-Acetyltransferase / metabolism*
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Cyanobacteria / genetics*
  • DNA Probes
  • Gene Expression Regulation, Bacterial / physiology*
  • Genetic Vectors*
  • Genome, Bacterial
  • Promoter Regions, Genetic*
  • Spectrometry, Fluorescence

Substances

  • DNA Probes
  • Chloramphenicol O-Acetyltransferase