TABLE 3.

Characteristics of M. avium subsp. paratuberculosis MBP fusion proteins

ProteinCalculated mol size (Da)aAmino acidsbAntigenic indexcYieldd (mg/ml)
MBP-gene 25021,434199/199Hydrophilic0.2
MBP-gene 25119,500179/179Hydrophilic0.7
MBP-gene 2529,68796/96Hydrophobic2.0
MBP-gene 2537,88174/7450/500.9
MBP-gene 25416,262146/14150/501.3
MBP-gene 25525,851241/241Hydrophilic0.3
MBP-gene 25615,120141/141Hydrophilic0.1
MBP-gene 2578,83787/87Hydrophilic1.0
MBP-gene 24021,734194/163Hydrophobic0.4
MBP-gene 24130,724280/280Hydrophilic0.4
MBP-gene 22840,817369/216Hydrophilic0.6
MBP-gene 21711,567106/10650/502.1
MBP-gene 218-C91,530835/365Hydrophilic1.4
MBP-gene 218-N91,530835/353Hydrophilic1.8
MBP-gene 21910,00487/85Hydrophilic0.04
MBP-gene 15920,655185/185Hydrophobic0.2
MBP-gene 13517,018157/141Hydrophobic1.0
MBP-gene 5621,116193/193Hydrophilic0.7
MBP-gene 5725,485252/25250/501.4
MBP-gene 3818,730173/170Hydrophilic0.06
MBP-gene 1036,380322/205Hydrophilic0.8
MBP-gene 1121,826191/191Hydrophilic0.7
  • a Represents the calculated size of the entire predicted coding sequence but does not represent the size of the fusion protein.

  • b Total number of amino acids in predicted coding sequence/number of amino acids represented in the MBP fusion. In some instances, the mycobacterial protein of interest was truncated to minimize toxicity and/or enhance expression in E. coli.

  • c The hydrophilic nature of a protein is considered a crude estimate of the protein's antigenicity. M. paratuberculosis proteins were analyzed by Kyte-Doolittle hydropathy plots (20) using MacVector sequence analysis software. Proteins that were more than 60% hydrophilic were termed “hydrophilic,” proteins less than 40% hydrophilic were termed “hydrophobic,” and proteins displaying 40 to 60% hydrophilicity were termed “50/50.”

  • d The purification yield was determined by using a Bio-Rad protein assay on pooled fractions.