TABLE 1.
Characteristics of strains, phages, plasmids, and primers used in this study
| Strain, phage, plasmid, primer, peptide | Characteristics, reference, and/or sourcea |
|---|---|
| Strains | |
| M. avium subsp. paratuberculosis strain 6783 | Clinical isolate, DSM 44135 |
| 14 M. avium subsp. paratuberculosis bovine clinical isolates | Fecal and lymph node isolates from clinically diseased cattle from different herds in northern Germany |
| M. avium subsp. avium strain ATCC 25291 | DSM 44156 |
| M. smegmatis mc2155 | 42 |
| M. scrofulaceum | NCTC 10803 |
| M. bovis BCG | NCTC 5692 |
| M. bovis | NCTC 10772 |
| M. kansasii | NCTC 10268 |
| M. microti | NCTC 8710 |
| M. gordonae | NCTC 10267 |
| M. fortuitum | NCTC 10394 |
| E. coli DH5αF′ | F′/endA1 hsdR17 (rK−mK+) supE44 thi-1 recA1 gyrA (Nalr) relA1 Δ(lacZYA-argF) U169 deoR [φ80dlacΔ(lacZ)M15] (35) |
| Phage fMptD | Phage isolated from the Ph.D.-12 phage display library with the specific sequence 5′-GGG AAG AAT CAT CAT CAG CAT CAT AGG CCT CAG-3′ |
| Plasmids | |
| pUC19 | E. coli cloning vector carrying an ampicillin resistance determinant (Pharmacia) |
| pGEX5x-3 | E. coli expression vector for the construction of glutathione S-transferase fusion proteins (Pharmacia) |
| pMV361 | Integrative mycobacterial shuttle vector carrying a kanamycin resistance determinant (9) |
| pRDIII300 | Fragment RDIII300 in pUC19 (this work) |
| pRDIII301 | Fragment RDIII301 in pUC19 (this work) |
| pRDIII302 | 4-kb DpnII fragment in pGH432 obtained by hybridization with a probe derived from the 5′ end of RDIII300 (this work) |
| pRDIII303 | 4-kb DpnII fragment in pGH432 obtained by hybridization with a probe derived from the 3′ end of RDIII300 (this work) |
| pRDIII320 | Fragment RDIII300 in pMV361 (Fig. 1, inset) (this work) |
| pRDIII320Δ1 | Deletion derivative of pRDIII320 with a 3,201-bp FseI fragment deleted (Fig. 1, inset) (this work) |
| pRDIII320Δ2 | Deletion derivative of pRDIII320 with a 2,410-bp SmaI-SacI fragment deleted (Fig. 1, inset) (this work) |
| pRDIII350 | PCR fragment obtained with primers ABC3 and ABC4, cut with BamHI and EcoRI, and ligated into pGEX5x-3 |
| Primers | |
| RBam12 | 5′-GAT CCT CGG TGA-3′ (29) |
| RBam24 | 5′-AGC ACT CTC CAG CCT CTC ACC GAG-3′ (29) |
| IPIII30 | 5′-CTA TGC GCA CTG ACG CTT C-3′ (this work) |
| IPIII31 | 5′-TTC CGA AGA ATC CGA TGA G-3′ (this work) |
| ABC3 | 5′-CCG CGG ATC CGC TTA CGA CGG AGG TCA A-3′ (this work) |
| ABC4 | 5′-GCC GGA ATT CGA TGT TGA TGA GAAT CCC T-3′ (this work) |
| ISMav1 | 5′-GTA TCA GGC CGT GAT GGC GG-3′ |
| ISMav2 | 5′-CGC CAC CAG CGC TCG ATA CA-3′ |
| Peptides | |
| aMptD | Biotin-aminohexacarbonic acid-GKNHHHQHHRPQ |
| aMPr | Biotin-aminohexacarbonic acid-HSQPKQVKKASR (control peptide) |
a
The nucleotides in boldface indicate the restriction enzyme sites used for cloning into pGEX5x-3. DSM and NCTC strains were from the Deutsche Gesellschaft für Mikroorgansismen und Zellkulturen (Braunschweig, Germany) and the National Collection of Type Cultures (Colindale, United Kingdom), respectively.