Article Figures & Data
Figures
- Fig. 1.
Comparison of the deduced PhoP amino acid sequences ofY. pestis, Y. pseudotuberculosis (Y. ptb), Y. enterocolitica (Y. ent), E. coli, and Salmonella serovar Typhimurium (S. typhm), aligned by using Clustal V multiple sequence alignment software. Identical amino acids (●), conserved changes (‖), and the essential aspartic acid residue (∗) are shown.
- Fig. 2.
Survival of Y. pestis wild-type (GB) andphoP mutant (SAI2.2) strains after uptake by J774 macrophages. Results shown are the means of duplicate determinations in two separate experiments, with standard errors.
- Fig. 3.
2D gel electrophoresis protein expression profiles for the Y. pestis wild-type strain (a) and Y. pestis phoP mutant strain (b) grown at 28°C. Differences in protein expression are highlighted (0). Mr, molecular weights in thousands.
- Fig. 4.
2D gel electrophoresis protein expression profiles for the Y. pestis wild-type strain (A) and Y. pestis phoP mutant (B) grown at 37°C. Differences in protein expression are highlighted (0). Mr, molecular weight in thousands.
Tables
- Table 1.
Bacterial strains and plasmids used in this studya
Strain or plasmid Relevant characteristic(s) Source or reference Strains Y. pestis GB Virulent wild-type strain, bv. orientalis 33 SAI2.2 Y. pestis GB phoPmutant This study Y. pseudotuberculosis YPIII pIB1 Virulent wild-type strain, serotype III 5 Y. enterocolitica 8081 Virulent wild-type strain, serotype O8 5 E. coli XL2-Blue MRF′ Cloning strain Stratagene CC118 λpir Cloning strain 29 S17 λpir pNJ5000 Triple mating strain with Tetr helper plasmid 18 Plasmids pUC19 Apr Pharmacia pCVD442 Apr Sucs, suicide vector 9 pNJ5000 Tetr, for triple mating 18 pYP5 pUC19 containing 262-bp PCRDOP gene fragment of Y. pestis phoP This study pYPTB7 pUC19 containing 262-bp PCRDOP gene fragment ofY. pseudotuberculosis phoP This study pDA5 pUC19 plus 4-kb DraI chromosomal fragment containingphoP This study pSA2 pUC19 and 1.5-kbSspI chromosomal fragment containing phoP This study pSAI1 pSA2 with 31-bp deletion inphoP This study pSAI2 Donor plasmid for conjugation; Apr Sucs, pSA1XbaI-PvuII fragment containing the mutatedphoP gene cloned into XbaI-SmaI pCVD442 This study ↵a Abbreviations: Apr, ampicillin resistant; Tetr, tetracycline resistant; Sucs, sucrose sensitive.
- Table 2.
Oligonucleotides used for PCR
Primer PCR method Strand Sequence (5′–3′)a, b P3 PCRDOP + AATCTGCAGYTNMGNCAYCAYYTNAANGT P4 PCRDOP − CCAAGCTTNARNACYTCNACYTTPTCYTG P9 IPCRM + CCAGATCTGGATGGCTTAAGCCTTATC P10 IPCRM − AAAGATCTTATCTGGGCCATGTTCCTG P24 phoP specific − ACTTTATCTTGCCAGCTTT P27 phoP specific + CGCGTTGTTGCGTCACCAT P41 phoPamplification + ACCTATCACCAGATATTGGCGTG P42 phoPamplification − CCCATCATCATTCTACTGATGTGCG
