Recombinant porin of Haemophilus influenzae type b.
A protein of approximately 40 kDa in the outer membrane of Haemophilus influenzae type b (Hib) behaves as a porin and permits transmembrane diffusion of low-molecular-weight solutes. The gene for Hib porin was cloned from an M13 library of chromosomal DNA of Hib strain ATCC9795. The gene was subcloned into a new transfer vector as a prerequisite for its use in the baculovirus expression vector system. Pure recombinant virus (AcPORIN) was isolated. On infection of a cultivated insect cell line Sf9 with AcPORIN, a novel protein was detected in cell lysates, and this novel protein reacted with an anti-Hib porin monoclonal antibody. The purified recombinant Hib porin was tested for its pore-forming properties in a synthetic black lipid membrane. The biophysical activity of purified recombinant Hib porin was identical to porin isolated from the bacterial outer membrane.
Coulton
J W
JW
Department of Microbiology and Immunology, McGill University, Montreal, Canada.
Chin
A C
AC
Vachon
V
V
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D016367:Baculoviridae / Q000235:genetics; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D006193:Haemophilus influenzae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D007473:Ion Channels / Q000235:genetics*; D018272:Porins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D001425:Bacterial Outer Membrane Proteins; D007473:Ion Channels; D018272:Porins; D011994:Recombinant Proteins