Preparation of luciferase-bacterial magnetic particle complex by artificial integration of MagA-luciferase fusion protein into the bacterial magnetic particle membrane.
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.
Matsunaga
Tadashi
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan. tmatsuna@cc.tuat.ac.jp
Arakaki
Atsushi
A
Takahoko
Mikako
M
D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000096:biosynthesis*; D001709:Biotechnology / Q000379:methods*; D002021:Buffers; D027682:Cation Transport Proteins / Q000096:biosynthesis*; D004926:Escherichia coli; D006531:HEPES; D008156:Luciferases / Q000302:isolation & purification*; D008163:Luminescent Measurements; D008280:Magnetics; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology*; D013010:Sonication
D001426:Bacterial Proteins; D002021:Buffers; D027682:Cation Transport Proteins; C097085:MagA protein, Magnetospirillum; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D006531:HEPES