Preparation of luciferase-bacterial magnetic particle complex by artificial integration of MagA-luciferase fusion protein into the bacterial magnetic particle membrane

Biotechnol Bioeng. 2002 Mar 20;77(6):614-8. doi: 10.1002/bit.10114.

Abstract

MagA is an iron-translocating protein found in the membranes of magnetic bacterium. Luciferase-bacterial magnetic particle (BMP) complexes were prepared by artificially inserting MagA-luciferase fusion proteins into the membranes of BMPs from Magnetospirillum magneticum strain AMB-1. Fusion proteins were from recombinant Escherichia coli membranes. MagA-Luc fusion proteins were integrated by sonication in vitro. Successful integration of fusion proteins was confirmed by luciferase luminescence on BMPs. Maximum luminescence was obtained after sonication for 3 min with a solution containing 300 mM NaCl, and is 18 times higher compared with recombinant Luc-BMPs generated using previously reported gene fusion techniques.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Artificial Gene Fusion
  • Bacterial Proteins / biosynthesis*
  • Biotechnology / methods*
  • Buffers
  • Cation Transport Proteins / biosynthesis*
  • Escherichia coli
  • HEPES
  • Luciferases / isolation & purification*
  • Luminescent Measurements
  • Magnetics*
  • Membrane Fusion*
  • Membrane Proteins / chemistry
  • Membrane Proteins / physiology*
  • Recombinant Fusion Proteins / physiology*
  • Sonication

Substances

  • Bacterial Proteins
  • Buffers
  • Cation Transport Proteins
  • MagA protein, Magnetospirillum
  • Membrane Proteins
  • Recombinant Fusion Proteins
  • Luciferases
  • HEPES