Four strains of the swine pathogen Actinobacillus pleuropneumoniae, namely, the type strain (ATCC 27088), the "reference" strain of biotype 2 (Bertschinger 2008/76) and two additional biotype 1 strains, strain BC181, which is less virulent than the type strain, and strain K17 (reference strain of serotype 5A), which was isolated from a lamb, were investigated with respect to iron acquisition. All four strains produced iron-repressible outer membrane proteins. However, only strains ATCC 27088 and Bertschinger 2008/76 could acquire iron from porcine transferrin. No organism could utilize human, bovine or ovine transferrin, or ovine or porcine lactoferrin. Haemoglobin supported good growth of all strains except K17 (which also failed to acquire iron from haemin). In all cases, iron acquisition from transferrin or haemoglobin required direct contact between the organisms and the proteins. Total membranes derived from iron-restricted organisms were subjected to an affinity isolation technique based on biotinylated porcine transferrin and streptavidin-agarose, and the following polypeptides were isolated: 99 kDa and 64 kDa from strain ATCC 27088; 93 kDa from strain Bertschinger 2008/76; 95 kDa (trace amounts) and 60 kDa from strain BC181; none from strain K17. These polypeptides appear to be transferrin receptor components. The 99 kDa polypeptide (TBPl) from the type strain was purified by SDS-PAGE and transferred electrophoretically onto polyvinylidene difluoride membrane. The N-terminal amino acid sequence of the polypeptide was determined commercially. A commercially-synthesized oligonucleotide probe was used to clone the gene encoding the TBPl of the type strain in competent Escherichia coli DH5$ alpha$ cells.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.28720 |
| Date | January 1995 |
| Creators | D'Silva, Colin Gerard |
| Contributors | Niven, Donald F. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Natural Resource Sciences.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001459930, proquestno: NN05692, Theses scanned by UMI/ProQuest. |
Page generated in 0.0017 seconds