SUMMARY: Haemophilus somnus is a Gram-negative bacterial bovine pathogen which can cause disease or be carried asymptomatically. We previously showed that four serum-sensitive isolates from asymptomatic carriers lacked a 13.4 kb sequence of chromosomal DNA that was present in two virulent serum-resistant strains. We have since sequenced 5 kb of the 13.4 kb fragment from a serum-resistant strain, which contained an open reading frame (ORF) of at least 4.5 kb. From Western blot analysis, the ORF was shown to encode a 76 kDa protein (p76) that co-migrated with a 76 kDa H. somnus surface protein. Both the recombinant and natural p76 reacted with convalescent-phase serum from a cow in an experimental H. somnus abortion study. The translational start site for p76 was identified by deletion analysis of subclones of the 5 kb cloned sequence. The 4.5 kb ORF contained 1.2 kb tandem direct repeats (DRs), with 65% identity between the two repeats at the protein level. The 5' DR (DR1) included the start site for the 76 kDa protein, and DR2 had a flanking inverted repeat, suggestive of an insertion-sequence-like element.
BailieW.E.,
ColesE.H.,
WeideK.D.1973; Deoxyribonucleic acid characterization of a microorganism isolated from infectious thromboembolic meningoencephalomyelitis of cattle.. International Journal of Systematic Bacteriology 23:231–237
BarbourA.G.,
CarterC.J.,
BurmanN.,
FreitagC.S.,
GaronC.F.,
BergstromS.1991; Tandem insertion sequence-like elements define the expression site for variable antigen genes of Borrelia hermsii. . Infection and Immunity 59:390–397
ColeS.P.,
GuineyD.G.,
CorbeilL.B.1992; Two linked genes for outer membrane proteins are absent in four non-disease strains of Haemophilus somnus. . Molecular Microbiology 6:1895–1902
CorbeilL.B.,
BlauK.,
PrieurD.J.,
WardA.C.S.1985; Serum susceptibility of Haemophilus somnus from bovine clinical cases and carriers.. Journal of Clinical Microbiology 22:192–198
DunnJ.J,
StudierF.W.1983; Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements.. Journal of Molecular Biology 166:477–535
EisenbergS.,
FinerM.1980; Cleavage and circularization of single-stranded DNA: a novel enzymatic activity of ɸX174 A* protein.. Nucleic Acids Research 22:5305–5315
GilmoreR.D.JRJosteN.,
McdonaldG.A.1989; Cloning, expression and sequence analysis of the gene encoding the 120 kDa surface-exposed protein of Rickettsia rickettsii. . Molecular Microbiology 3:1579–1586
GilmoreR.D.JRCieplakN.JRPolicastroP.F.,
HackstadtT.1991; The 120 kilodalton outer membrane protein (rOmpB) of Rickettsia rickettsii is encoded by an unusually long open reading frame: evidence for protein processing from a large precursor.. Molecular Microbiology 5:2361–2370
GogolewskiR.P.,
KaniaS.A.,
InzanaT.J.,
WiddersP.R.,
LiggittH.D.,
CorbeilL.B.1987; Protective ability and specificity of convalescent serum from calves with Haemophilus somnus pneumonia.. Infection and Immunity 55:1403–1411
GroomS.C.,
LittleP.B.,
RosendalS.1988; Virulence differences among three strains of Haemophilus somnus following intratracheal inoculation of calves.. Canadian Journal of Veterinary Research 52:349–354
HumphreyJ.D.1982Haemophilus somnus: colonization of the bovine reproductive tract. [As quoted in Groom et al. (1988) and Humphrey & Stephens (1983).] PhD thesis University of Guelph, Ont., Canada.:
InzanaT.J.,
CorbeilL.B.
1987; Further characterization of the growth requirements of Haemophilus somnus : development of a defined medium.. American Journal of Veterinary Research 48:366–369
InzanaT.J.
1992; Immune response of cattle to Haemophilus somnus lipid A-protein conjugate vaccine and efficacy in a mouse abortion model.. American Journal of Veterinary Research 53:175–179
IsbergR.R.,
LaazaarA.L.,
SyvanenM.1982; Regulation of Tn5 by the right-repeat proteins: control at the level of the transposition reaction.. Cell 30:883–892
JohnsonR.C.,
ReznikoffW.S.1984; Role of the IS50R proteins in the promotion and control of Tn5 transposition.. Journal of Molecular Biology 177:645–661
MillsJ.A.,
VenkatesanM.M.,
BaronL.S.,
BuysseJ.M.1992; Spontaneous insertion of an ISi-like element into the virF gene is responsible for avirulence in opaque colonial variants of Shigella ftexneri 2a.. Infection and Immunity 60:175–182
NotarnicolaS.M.,
McintoshM.A.
1990; Multiple translational products from a Mycoplasma hyorhinis gene expressed in Escherichia coli. . Journal of Bacteriology 172:2986–2995
PritchardA.E.,
VasilM.L.1990; Possible insertion sequences in a mosaic genome organization upstream of the exotoxin A gene in Pseudomonas aeruginosa. . Journal of Bacteriology 172:2020–2028
RossT.K.,
AchbergerE.C.,
BraymerH.D.1989; Nucleotide sequence of the McrB region of Escherichia coli K-12 and evidence for two independent translational initiation sites at the mcrB locus.. Journal of Bacteriology 171:1974–1981
SharpP.M.,
CoweE.,
HigginsD.G.,
ShieldsD.C.,
WolfeK.H.,
WrightF.1988; Codon usage patterns in Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Drosophila melanogaster and Homo sapiens; a review of the considerable within-species diversity.. Nucleic Acids Research 16:8207–8211
TheisenM.,
PotterA. A.
1992; Cloning, sequencing, expression, and functional studies of a 15000-molecular-weight Haemophilus somnus antigen similar to Escherichia coli ribosomal protein S9.. Journal of Bacteriology 174:17–23
TheisenM.,
RiouxC.R.,
PotterA.A.1992; Molecular cloning, nucleotide sequence, and characterization of a 40000-molecular-weight lipoprotein of Haemophilus somnus. . Infection and Immunity 60:826–831
WiddersP.R.,
PaisleyL.G.,
GogolewskiR.P.,
EvermannJ.F.,
SmithJ.W.,
CorbeilL.B.1986; Experimental abortion and the systemic immune response in cattle to Haemophilus somnus. . Infection and Immunity 54:555–560
WiddersP.R.,
DorranceL.A.,
YarnallM.,
CorbeilL.B.1989; Immunoglobulin-binding activity among pathogenic and carrier isolates of Haemophilus somnus. . Infection and Immunity 57:639–642
YarnallM.,
WiddersP. R.,
CorbeilL.B.1988b; Isolation and characterization of Fc receptors from Haemophilus somnus. . Scandinavian Journal of Immunology 28:129–137