Recombinant Porin of Haemophilus influenzae Type b

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Abstract
A protein of ∼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 ofHib strain ATCC 9795. 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.

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