Table 1.
Selected ORFs of M. hyopneumoniae and primer sequences
| Recombinant protein annotationa | Function | Selected fragment (aa) | Primer sequencec |
|---|---|---|---|
| MHP0107 | Protein P102 (copy 2) | 646–948 | 5′-CACCGGATCCGTATCAAAAGCAGATCGA-3′ (F) |
| 5′-CCCAAGCTTTTATTGTTTTATATAATTACTAAT-3′ (R) | |||
| MHP0272 | Protein P97-paralog | 228–708 | 5′-CACCGGATCCAAAATAAAAGAAAAATTGTTT-3′ (F) |
| 5′-CCGGTACCTCAAGCCCGAACTTT-3′ (R) | |||
| MHP0418 | Hypothetical protein | 145–500 | 5′-CACCGGATCCGAAAATTTAGCGCCATAT-3′ (F) |
| 5′-CCCAAGCTTTCATTTTTTAACTGAAATTGG-3′ (R) | |||
| 5′-AGGCATTGgTATTCTTTATTAG-3′ (FM) | |||
| 5′-AGAATAcCAATGCCTTTCAG-3′ (RM) | |||
| MHP0443 | Hypothetical protein | 32–510 | 5′-CACCGGGTACCGAATGATTTTGTCTTTTTG-3′ (F) |
| 5′-CCCAAGCTTTCAGATTCTACCATTAATCTTT-3′ (R) | |||
| MHP0487 | Putative Mg2+ transporter | 1–280 | 5′-CACCGGATCCGATAAAAAAAGTTCTTTTCTTTT-3′ (F) |
| 5′-CCCAAGCTTTCAAAAAACTCTTGATTTTACGAT-3′ (R) | |||
| MHP0660 | Hypothetical protein | 1–394 | 5′-CACCGGATCCATAAAAAAATCATCAAAAAT-3′ (F) |
| 5′-GGGGTACCTCACAGCCTTAGATTTAAA-3′ (R) | |||
| MHP0372b | Lppt lipoprotein | 497–883 | 5′-CACCTTTGATTTTCTTAATTTTTCAACA-3′ (F) |
| 5′-TTATTCCATATATTCGCTAAGGA-3′ (R) | |||
| 5′-TTCCTTGATTGgCGATCAAATATA-3′ (FM) | |||
| 5′-TGATCGcCAATCAAGGAATAAATC-3′ (RM) |
a
GenBank annotation of M. hyopneumoniae 7448 (NC_007332).
b
Directionally cloned into a Champion pET200D/TOPO His-tagged expression vector (Invitrogen).
c
F, forward primer; R, reverse primer; FM and RM, forward and reverse primers for site-directed mutagenesis of MHP0418 and MHP0372, respectively. Mutated nucleotides are shown in bold lowercase letters. Underlined nucleotides represent restriction sites of BamHI (GGATCC), HindIII (AAGCTT), and KpnI (GGTACC) enzymes.