The bacterium Moraxella lacunata is a causative agent of human conjunctivitis and keratitis. We have previously reported construction of plasmid pMxLl, which includes a 5.9-kb fragment on which the pilin gene inversion region of M. lacunata resides. The inversion region of pMxLl was shown to invert when pMxLl was in an Escherichia coli host cell. In this report, we present Western immunoblot analysis using Moraxelia bovis Epp63 anti-I and anti-Q pilin sera which demonstrate that pMxLl makes pilin only when in orientation 1. The type 4 (MePhe) class of bacterial pili is found on a wide variety of gram-negative bacteria (4), including Moraxella bovis (14) and Moraxella lacunata (12). The type 4 piliated (P+) bacteria of the family Neisseriaceae (including Moraxella, Branhamella, Neisseria, and other genera) display several phenotypic differences from the equivalent nonpiliated (P-) bacteria. These phenotypes include colony morphology, pitting of agar, autoagglutination, pellicle formation, hemagglutination, twitching motility, and increased competence for natural DNA transformation (for a review, see reference 10).We have been studying the type 4 pilin gene regions of M. bovis and M. lacunata. These organisms are very closely related, as determined by both DNA-DNA hybridization analysis (20) and transformation studies (2). M. bovis is the primary cause of bovine infectious keratoconjunctivitis (8), and M. lacunata is a pathogen which occasionally causes conjunctivitis in humans (3,16,19). A single strain of M. bovis is capable of producing one or the other of two pilin types, named Q (previously P) and I (previously x) for strain Epp63 (14,17). The switch in expression between the tfpQ and tfpl pilin genes is due to an inversion of a 2.1-kb region of DNA (5, 13). When tfpQ is in the expression locus it is in orientation 1, while orientation 2 has tfpI in the expression locus. We have shown that a gene (piv) adjacent to the inversion region is required for inversions to occur (12). We have described both similarities and differences between the * Corresponding author.