Specific immobilization of firefly luciferase through a biotin carboxyl carrier protein domain.
Firefly luciferase (Photinus pyralis) was fused with a histidine tag and a biotin carboxyl carrier protein (BCCP) domain at its amino terminus. Highly purified recombinant luciferase was obtained by a one-step purification protocol, utilizing immobilized metal affinity chromatography. The novel BCCP-luciferase had properties, stability, and activity similar to those of native luciferase. The biotin molecule on the BCCP domain allowed specific immobilization of BCCP-luciferase on avidin-coated surfaces via the biotin-avidin interaction.
Wang
C Y
CY
Department of Bioengineering, University of Utah, 2480 MEB, Salt Lake City, Utah, 84112, USA.
Hitz
S
S
Andrade
J D
JD
Stewart
R J
RJ
D000103:Acetyl-CoA Carboxylase; D000818:Animals; D001360:Avidin; D002352:Carrier Proteins; D001517:Coleoptera; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D054889:Fatty Acid Synthase, Type II; D008156:Luciferases
D002352:Carrier Proteins; D004800:Enzymes, Immobilized; D001360:Avidin; D008156:Luciferases; D054889:Fatty Acid Synthase, Type II; D000103:Acetyl-CoA Carboxylase; C079633:biotin carboxyl carrier protein