Article Figures & Data
Figures
- Fig. 1.
Genes identified by immunoscreening of Mycoplasma haemofelis Lambda ZapII libraries. The clones discovered are indicated by the abbreviations listed in Table 1, and the location in the genome of M. haemofelis is shown. The 16S rRNA gene and the gene coding for DnaA (chromosomal replication initiator) are marked in gray as reference points. The scale is in kilobases.
- Fig. 2.
SDS-PAGE and Western blot analyses of the putative antigens P6D-orf0908, P10C-orf1238, P10E-orf0279, P10C-orf1239, P21B-orf1546l, and P29-orf0177. (Top) SDS-PAGE showing the expression of putative antigens (SimplyBlue SafeStain; Invitrogen); (middle) Western blot results of the putative antigens against the His6 tag; (bottom) Western blot results of the putative antigens against convalescent pooled plasma from an experimentally infected cat. Lane 1, fragment of the DnaK control (approximately 31.75 kDa); lane 2, fragment of P6D-orf0908 (approximately 18.65 kDa); lane 3, fragment of P10C-orf1238 (approximately 20.52 kDa); lane 4, fragment of P10E-orf0279 (approximately 19.07 kDa); lane 5, fragment of P10C-orf1239 (approximately 18.88 kDa); lane 6, fragment of P21B-orf1546 (approximately 12.56 kDa); lane 7, fragment of P29-orf0177 (approximately 21.71 kDa); lanes M, molecular mass markers (Precision Plus Protein Kaleidoscope; Bio-Rad).
Tables
- Table 1.
Reactive clones by immunoscreening of Mycoplasma haemofelis Lambda ZapII libraries and BLASTx results against nonredundant protein sequence database
Insert identification GenBank accession no. Sequence similarity by BLASTx against the Mollicutes E value P3-orf1165a,b GS928052 ACQ84443.1, adhesin (Mycoplasma hyopneumoniae) 0.002 P5-orf1816a,b,c GS928053 NP_853366.1, thymidine phosphorylase (M. gallisepticum) 0.37 P6D-orf0908a,b,d GS928054 ZP_06610317.1, hypothetical protein MALL_0643 (M. alligatoris) 0.44 P6D-orf0909b GS928055 ZP_06610593.1, hypothetical protein MALL_0515 (M. alligatoris) 0.63 P7-orf0259 GS928056 ACU78513.1, triose-phosphate isomerase (M. mycoides) 8E-36 P7-orf0260 GS928057 NP_073101.2, phosphoglyceromutase (M. genitalium) 9E-128 P7-orf0261 GS928058 BAH70152.1, hypothetical protein (M. fermentans) 0.026 P7-orf0262a GS928059 NSe P7-orf0263a GS928060 ABD47695.1, adhesin-like protein P146 (M. hyopneumoniae) 1.4 P7-orf0264 GS928061 AAZ44718.2, conserved hypothetical protein (M. hyopneumoniae) 0.22 P9D-orf1202 GS928062 ZP_06610215.1, type I restriction modification DNA protein (M. alligatoris) 2E-15 P9D-orf1203 GS928063 YP_003303059.1, type I restriction enzyme specificity protein (M. hominis) 0.005 P9D-orf1204 GS928064 ZP_02931536.1, type I restriction enzyme S protein (Ureaplasma parvum) 4E-10 P9D-orf1205 GS928065 YP_003303059.1, type I restriction enzyme specificity protein (M. hominis) 2E-12 P10B-orf1747a,b GS928066 YP_002284694.1, putative lipoprotein (Ureaplasma urealyticum) 3.1 P10B-orf1748 GS928067 YP_003515875.1, hypothetical protein MAGa7180 (M. agalactiae) 0.17 P10B-orf1749a,b GS928068 NS 1.1 P10B-orf1750 GS928069 YP_003560289.1, hyaluronoglucosaminidase (M. crocodyli) 1.1 P10C-orf1238a,b,d GS928070 YP_002000188.1, massive surface protein MspK (M. arthritidis) 0.004 P10C-orf1239a,b,d GS928071 ZP_04563878.1, transcriptional regulator (Mollicutes bacterium D7) 0.002 P10E-orf0279a,d GS928072 NP_757929.1, hypothetical protein MYPE5440 (M. penetrans) 3E-50 P10E-orf0280 GS928073 NP_975564.1, pseudouridylate synthase D (M. mycoides) 1E-41 P15-orf0941 GS928074 YP_002960937.1, hypothetical protein MCJ_004270 (M. conjunctivae) 0.0024 P15-orf0942 GS928075 NS P15-orf0943b GS928076 ACU78785.1, conserved hypothetical protein (M. mycoides) 0.28 P15-orf0944 GS928077 AAO39838.1, AvgC variable lipoprotein (M. agalactiae) 0.17 P15-orf0945a,b GS928078 YP_002000023.1, massive surface protein MspF (M. arthritidis) 0.63 P15-orf0946a,b GS928079 ZP_02695921.2, hypothetical protein UUR13 (U. urealyticum) 1.8 P15-orf0947a,b GS928080 ZP_06610731.1, conserved hypothetical protein (M. alligatoris) 0.37 P15-orf0948b GS928081 YP_279005.1, lysyl-tRNA synthetase (M. hyopneumoniae) 0.63 P15-orf0949b GS928082 NP_758309.1, phenylalanyl-tRNA synthetase subunit beta (M. penetrans) 0.48 P17A-orf1526 GS928083 CAB62239.1, P75 protein (M. hominis) 0.28 P17A-orf1527a,b GS928084 ZP_04564868.1, conserved hypothetical protein (Mollicutes bacterium D7) 0.002 P17A-orf1528b GS928085 ZP_02971377.1, conserved hypothetical protein (U. parvum) 0.37 P17A-orf1529b GS928086 YP_016078.1, hypothetical protein MMOB3810 (M. mobile) 0.37 P18-orf0127 GS928087 NP_757967.1, hypoxanthine-guanine phosphoribosyltransferase (M. penetrans) 6E-07 P18-orf0128a GS928088 YP_002000162.1, hypothetical protein MARTH (M. arthritidis) 0.13 P20/22-orf0326a GS928089 NP_757466.1, DNA-directed RNA polymerase subunit beta (M. penetrans) 0 P21A-orf0675a,b GS928090 NP_757933.1, translocase (Mycoplasma penetrans) 3.1 P21B-orf1544a,b GS928091 YP_002000128.1, massive surface protein MspH (M. arthritidis) 0.044 P21B-orf1545 GS928092 NP_757749.1, hypothetical protein MYPE3620 (M. penetrans) 0.057 P21B-orf1546a,b,d GS928093 YP_001256183.1, hypothetical protein MAG_0390 (M. agalactiae) 0.097 P21B-orf1547b GS928094 NP_975173.1, hypothetical protein MSC_0170 (M. mycoides) 1.1 P21B-orf1548b GS928095 YP_001799373.1, hypothetical protein PAa (C. Phytoplasma australiense) 0.13 P24-orf1679a GS928096 YP_002000015.1, massive surface protein MspC (M. arthritidis) 1.13 P24-orf1680 GS928097 NS P24-orf1681b GS928098 NP_758083.1, hypothetical protein MYPE6950 (M. penetrans) 0.28 P26-orf0285 GS928099 NP_853008.2, translation elongation factor Tu (EF-Tu) (M. gallisepticum) 8E-129 P26-orf0286 GS928100 NP_757969.1, adenylosuccinate synthetase (M. penetrans) 4E-110 P26-orf0287 GS928101 ADC31594.1, ribosomal biogenesis GTPase (M. gallisepticum) 2E-31 P27A-orf1350b GS928102 YP_002000022.1, massive surface protein MspE (M. arthritidis) 2E-03 P28-orf1521a,b GS928103 YP_002961131.1, hypothetical protein MCJ_006330 (M. conjunctivae) 0.130 P28-orf1522b GS928104 YP_016010.1, SWF/SNF family helicase (M. mobile) 0.630 P28-orf1523b GS928105 ZP_04563267.1, conserved hypothetical protein (Mollicutes bacterium D7) 0.220 P29-orf0175 GS928106 YP_001621362.1, ketose bisphosphate aldolase (Acholeplasma laidlawii) 5E-102 P29-orf0176 GS928107 NP_072863.1, cochaperone GrpE (M. genitalium) 4E-22 P29-orf0177a GS928108 NP_758284.1, heat shock protein DnaJ (M. penetrans) 2E-69 P29-orf0178 GS928109 NP_109915.1, elongation factor G (M. pneumoniae) 0 P32C-orf0088 GS928110 ZP_03079605.1, arginyl-tRNA synthetase (U. urealyticum) 7E-75 P33B-orf1097 GS928111 YP_002000188.1, massive surface protein MspK (M. arthritidis) 0.057 ↵a Putative proteins selected by bioinformatics analyses to be cloned and expressed by the Gateway system.
↵b Putative proteins predicted to have a transmembrane domain(s) by hidden Markov model and dense alignment surface methods.
↵c Shading indicates ORFs within the same plaque.
↵d Putative proteins that were reactive against immune plasma and negative against nonimmune plasma by Western blotting.
↵e NS, no significant similarity found.
