Table 1. Recombinant P. multocida proteins tested as vaccine candidates in chickens.
| Gene number a | Predicted molecular mass | Protein name and predicted function | Expression vector b | Solubility c | Signal P d | Immunogenic e | Reaction with P.m f | Protection |
| controls g | Killed whole cells | + | ||||||
| Adjuvant only | − | |||||||
| PM0040 | 81.2 | PfhR - putative outer membrane hemin receptor | pDEST-17 | I | + | + | − | − |
| PM0055 | 28.5 | acyl-CoA thioester hydrolase YfbB | pDEST-41BA | I | − | + | − | − |
| PM0056 | 54 | FhaC – Filamentous haemagglutinin two partner secretion protein | pDEST-17 | I | − | + | − | − |
| PM0076 | 74.5 | MapC protein | pDEST-17 | I | + | + | − | − |
| PM0098 | 41.9 | OapA - Cell envelope opacity-associated protein A | pBAD-DEST49 | I | − | + | − | − |
| PM0243 | 59.5 | Pm0243 - hypothetical protein | pDEST-41BA | S | + | + | + | − |
| PM0246 | 24.3 | LolB - outer membrane lipoprotein | pDEST-17 | I | + | + | − | − |
| PM0300 | 109.6 | HgbA - TonB-dependent haemoglobin receptor | pDEST-17 | I | + | + | − | − |
| PM0331 | 21.7 | OmpW - outer membrane protein | pDEST-17 | I | + | + | − | − |
| PM0337 | 113.2 | HgbB - Haemoglobin/haptoglobin binding protein | pDEST-17 | I | + | + | − | − |
| PM0355 | 29.9 | Pm0355 - hypothetical protein | pDEST-17 | I | − | + | − | − |
| PM0368 | 26.9 | Pm0368 - hypothetical protein | pDEST-41BA | S | + | + | − | − |
| PM0388 | 37.3 | OmpH1 - outer membrane porin | pDEST-17 | I | + | + | + | − |
| PM0389 | 38.6 | OmpH2 - outer membrane porin | pDEST-17 | I | + | + | − | − |
| PM0442 | 23.7 | Pm0442 - immunogenic membrane protein | pDEST-41BA | S | + | + | + | − |
| PM0519 | 12.5 | Pm0519 - hypothetical protein | pDEST-17 | I | − | + | − | − |
| PM0527 | 50.6 | TolC1 - outer membrane efflux channel | pDEST-17 | I | + | + | + | − |
| PM0542 | 75.1 | Glgx - glycogen operon protein | pDEST-41BA | I | − | + | − | − |
| PM0554 | 15.5 | PCP - peptidoglycan-associated lipoprotein cross reacting protein | pDEST-41BA | I | + | + | + | − |
| PM0586 | 29.9 | Plp4- lipoprotein | pDEST-17 | I | + | + | − | − |
| PM0612 | 11.4 | Pm0612 - Hypothetical outer membrane protein | pDEST-41BA | I | − | + | − | − |
| PM0618 | 99.4 | Pm0618 - aminopeptidase N | pDEST-41BA | I | − | + | − | − |
| PM0659 | 214.3 | Pm0659 - Large extracellular alpha-helical protein, lipoprotein | pDEST-17 | S | + | + | − | − |
| PM0680 | 77.4 | PrlC - oligopeptidase A | pDEST-41BA | I | − | + | + | − |
| PM0708 | 20.4 | Pm0708 - hypothetical outer membrane lipoprotein | pDEST-17 | I | + | + | − | − |
| PM0714-1 h | 91.4 | Hsf - hypothetical outer membrane protein | pDEST-17 | I | − | + | − | − |
| PM0714-2 h | 93.7 | Hsf - hypothetical outer membrane protein | pDEST-41BA | I | − | + | − | − |
| PM0714-3 h | 94.9 | Hsf - hypothetical outer membrane protein | pDEST-41BA | I | − | + | − | − |
| PM0741 | 89.4 | TonB dependent outer membrane haemoglobin/haemin binding receptor | pDEST-17 | I | + | + | − | − |
| PM0778 | 42.9 | HexD - putative polysaccharide export protein wza precursor | pDEST-41BA | S | + | + | + | − |
| PM0786 | 37.9 | OmpA - β-barrel membrane anchor protein | pDEST-17 | I | + | + | + | − |
| PM0803 | 90.3 | Pm0803 - iron regulated outer membrane protein | pDEST-17 | I | + | + | − | − |
| PM0881 | 18.4 | Pm0881 - hypothetical outer membrane protein | pDEST-17 | I | + | + | − | − |
| PM0892 | 17.1 | ImpA - hypothetical outer membrane protein | pDEST-41BA | I | − | + | − | − |
| PM0903 | 45.5 | Pm0903 - hypothetical outer membrane protein | pDEST-17 | I | + | + | − | − |
| PM0928 | 41.2 | MltA - murein transglycosylase A | pDEST-17 | I | + | + | − | − |
| PM0966 | 16.1 | Omp16 - Outer membrane protein P6 precursor; peptidoglycan-associated outer membrane lipoprotein | pDEST-17 | I | + | + | + | − |
| PM0979 | 14.5 | Pm0979 - immunogenic lipoprotein | pDEST-17 | I | + | + | − | − |
| PM0998 | 30.6 | Pm0998 - uncharacterised outer membrane protein | pDEST-41BA | I | + | + | + | − |
| PM1016 | 42.1 | Wza - putative polysaccharide export protein | pDEST-17 | I | + | + | + | − |
| PM1025 | 20.4 | Opa - outer membrane protein | pDEST-17 | I | + | + | + | − |
| PM1050 | 37.2 | Pm1050 - uncharacterised lipoprotein | pDEST-41BA | S | + | + | + | − |
| PM1064 | 30 | Pm1064 - uncharacterised lipoprotein | pBAD-DEST49 | I | + | + | − | − |
| PM1069 | 47.7 | Pm1069 - outer membrane protein P1 precursor | pDEST-41BA | I | + | + | + | − |
| PM1077 | 20.4 | Pm1077 - hypothetical protein | pDEST-41BA | S | + | + | + | − |
| PM1081 | 90.8 | Pm1081 - TonB dependent outer membrane haemoglobin/haemin binding receptor | pDEST-41BA | I | + | + | + | − |
| PM1225 | 49.1 | ComE - putative competence protein E | pDEST-17 | I | + | + | − | − |
| PM1238 | 36.9 | Pm1238 - putative O-sialoglycoprotein endopeptidase | pDEST-17 | I | − | + | + | − |
| PM1245 | 32.7 | Pm1245 - putative L-xylulose 5-phosphate 3-epimerase | pDEST-41BA | I | − | + | − | − |
| PM1426 | 35.4 | Phospholipase A - OMPLA | pDEST-41BA | I | + | + | + | − |
| PM1428 | 90.7 | Pm1428 - TonB dependent outer membrane haemoglobin/haemin binding receptor | pBAD-DEST49 | I | + | + | + | − |
| PM1451 | 30.4 | XynC - predicted esterase | pDEST-41BA | I | + | + | + | − |
| PM1501 | 27.4 | VacJ lipoprotein homolog, Surface lipoprotein H. influenzae | pDEST-17 | I | + | + | − | − |
| PM1517 | 37.3 | PlpE - protective outer membrane lipoprotein | pDEST-17 | I | + | + | + | + |
| PM1578 | 35.7 | Pm1578 - uncharacterised lipoprotein | pDEST-41BA | S | + | + | + | − |
| PM1611 | 19.6 | PM1611 - hypothetical protein | pDEST-17 | S | + | + | − | − |
| PM1614 | 49.6 | Pm1614 - outer membrane antigenic lipoprotein B | pDEST-17 | S | + | + | − | − |
| PM1622 | 95.8 | HasR - Ton B dependent haeme acquisition system receptor | pDEST-17 | I | + | + | + | − |
| PM1670 | 22.9 | RpiA - ribose-5-phosphate isomerase A | pDEST-41BA | S | − | + | − | − |
| PM1707 | 40.1 | PM1707 - uncharacterised protein | pDEST-41BA | S | + | + | + | − |
| PM1717 | 95.1 | PM1717 - uncharacterised protein | pBAD-DEST49 | I | + | + | − | − |
| PM1720 | 29.2 | Pm1720 - DNA uptake lipoprotein | pDEST-17 | I | + | + | + | − |
| PM1730 | 30.1 | MetQ - periplasmic methionine binding protein of ABC transporter | pDEST-41BA | I | + | + | + | − |
| PM1809 | 67 | Pm1809 - uncharacterised outer membrane protein | pDEST-17 | I | + | + | + | − |
| PM1826 | 25.5 | Pm1826 - uncharacterised lipoprotein | pDEST-41BA | I | + | + | + | − |
| PM1886 | 15.4 | Pm1886 - uncharacterised outer membrane protein | pDEST-17 | I | + | + | − | − |
| PM1898 | 22.4 | Pm1898 - uncharacterised lipoprotein | pDEST-41BA | S | − | + | − | − |
| PM1974 | 15.7 | Pm1974 - LysM domain/BON superfamily protein | pDEST-17 | I | − | + | + | − |
| PM1980 | 51.8 | TolC2 - outer membrane efflux channel | pDEST-41BA | I | + | + | − | − |
| PM1992 | 87.6 | Oma87– Omp85-like protein assembly protein | pDEST-17 | I | + | + | + | − |
| PM1993 | 21.3 | Skp - Skp export factor homolog | pDEST-17 | I | + | + | + | − |
| PM1979 | 69.3 | Pm1979 - peptidyl-prolyl cis-trans isomerase | pDEST-41BA | S | + | + | − | − |
| PM2008 | 20.8 | Pm2008 - putative fimbrial biogenesis and twitching motility protein | pDEST-17 | I | + | + | + | − |
a
- Gene numbers are from the Pm70 strain [28] homolog of the X-73 genes amplified and cloned.
b
- Expression vectors: pBAD-DEST49, a Gateway® destination vector that features a thioredoxin fusion tag; pDEST-41BA a modified Gateway-adapted expression vector containing a NusA solubility tag [20] (originally derived from pLIC-NusA [48]); pDEST-17, a Gateway® destination vector for production of tagless native protein.
c
- I = insoluble; S = soluble protein.
d
- SignalP server signal peptide prediction. In general PCR primers were designed so as to amplify the gene region encoding the mature length protein, excluding the signal sequence, except where a signal sequence could not be predicted; in this case the primers were designed to encompass the entire gene.
e
- immunogenic reaction against the recombinant protein.
f
- serum reacted with a protein of appropriate size in a whole cell lysate derived from P. multocida grown in vitro at 37°C.
g
- positive (X-73 whole killed cells) and negative controls used in the vaccine trial.
h
- Due to its large size, PM0714 was purified in three overlapping sub-fragments spanning the entire protein.