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United States Department of Agriculture

Agricultural Research Service

Title: Molecular Cloning, Expression, and DNA Sequence Analysis of a Gene Encodingan Outer Membrane Protein of Brachyspira (Serpulina) Pilosicoli

Authors
item Alt, David
item Trott, Darren
item Stanton, Thaddeus

Submitted to: Pig Veterinary Society International Congress Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: September 20, 2000
Publication Date: N/A

Technical Abstract: A monoclonal antibody, 2E10, prepared against outer membrane enriched vesicles from Brachyspira pilosicoli, strain 95-1000, was used to identify reactive clones from a genomic library of 95-1000. Two positive clones were sequenced and found to overlap, forming a 4.9 kb fragment. A putative 540 bp open reading frame within this sequence encodes a 19 kDa protein. Sequence analysis of the predicted protein showed homology at the N terminus to a previously reported 16 kDa lipoprotein, smpA, from B. hyodysenteriae. The remainder of the protein sequence did not show any homology with other proteins in Genbank. The protein was identified in Escherichia coli cell fractions using MAb 2E10 as a band of approximately 27 kDa. The band was most prominent in the outer membrane fractions of E. coli. N-terminal sequencing of the protein identified by MAb 2E10 confirmed the protein as that encoded by the 540 bp open reading frame.

Last Modified: 7/25/2014
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