Deficiency of the cytochrome P-450 steroid 21-hydroxylase (21-OHase) which causes Congenital Adrenal Hyperplasia (CAH) is a monogenic autosomal recessive disorder which is linked to HLA. There are two 21-OHase genes in man, A and B, and they are mapped to the HLA class III region ~ 3 kb 3' to the complement genes C4A and C4B, respectively. Two genes encoding 21-OHase were isolated, characterized and sequenced. Both 21-OHase genes are ~ 3.3 kb in length and are split into 10 exons by nine introns. Comparison of the two genes showed that although they are highly conserved, there are three deleterious mutations in the 21-OHase A gene which cause frameshifts and introduce in phase premature termination codons. Thus the 21-OHase A gene is a pseudogene. Comparison of the 21-OHase B gene to the other cytochrome P-450 sequences revealed that although the cysteine-429 was conserved in 21-OHase, there is very little homology with other cytochrome P-450, indicating it belongs to a separate family of genes within the superfamily. Clear evidence of polymorphism in 21-OHase is apparent on comparison with other 21-OHase B sequences. There is a size polymorphism of 494 and 495 amino acids. The differing severities of 21-OHase deficiency in CAH may be due to allelic variants of the 21-OHase B gene, since in most cases the defect is not due to gene deletion (Rumsby et al., 1986). A 21-OHase B gene from a patient with CAH was characterized and sequenced. There were 13 nucleotide alterations in his single 21-OHase B gene, one of which at codon 269 caused a serine to change to a threonine residue. The G → C transversion in the 21-OHase B gene from the patient at codon 269 introduced a new NcoI restriction site into the gene. This restriction fragment length polymorphism (RFLP) was used to study other patients with CAH and normal individuals. The NcoI RFLP was found not to be confined to the 21-OHase B gene but was also present in some 21-OHase A genes. It is likely therefore that the mutation occurred in the pseudogene first and then transferred to some 21-OHase B genes.

This thesis describes studies on the binding of peptides to the murine major histocompatibility complex (MHC) class I molecule H-2D^b (D^b). The expression of the recombinant soluble D^b molecule in Chinese hamster ovary cells and its subsequent purification by nickel affinity chromatography, gel filtration, and preparative native isoelectric focusing are reported. The product is the correct molecule, homogeneous, a dimer of dimers, and free of endogenous peptide. A novel binding assay based on the enhancement of natural tryptophan fluorescence by the binding of peptide is introduced. This assay is used to determine melting curves of the empty and peptide-loaded protein, and to measure association rate constants by stopped-flow fluorescence spectroscopy. Radioligand binding measurements of equilibrium as well as association and dissociation rate constants and their temperature dependence are reported. In agreement with earlier observations, the ratio of association and dissociation rate constants is much larger than the equilibrium association constant. Fluorescence anisotropy decay spectroscopy gives evidence for conformational alterations in the D^b molecule upon peptide binding. The data, possible errors and ways to avoid them, and mathematical models of binding are discussed to obtain an overall picture of the binding process.

Both major histocompatibility genes (MHC) and infection have been shown to influence urinary odours in mice, and mice may use MHC-associated odours to detect kin for the purpose of choosing communal nesting partners, or to avoid mating with close relatives. Additionally, mice may use infection-related odours to avoid mating with sick individuals or those genetically susceptible to infection. I tested the above theories using urine as the source of odour. In a series of odour preference experiments, employing two MHC-congenic strains of mice (B10 and B10.Q), I tested the proposition that females prefer MHC-similar female odours when choosing female relatives as nesting partners, and that females prefer MHC-disparate male odours for the purpose of disassoratative mating. I found that females did not display a preference for MHC-similar female odours or MHC-disparate male odours. However, when B10.Q male urine donors were infected with 100 L3 larvae of Heligmosomoides polygyrus (Nematoda), both strains of female displayed a significant preference for the odours of uninfected B10 males. To determine whether female odour preference for uninfected males extended to actual mate choice, I subsequently examined female mate choice in a controlled setting. Using CD-1 outbred mice, I found that females preferred to mate with uninfected males over males subclinically infected with 200 L3 of H. polygyrus , as evidenced by first ejaculation preferences. In this experiment, males were tethered, and as such, male dominance interactions were prevented. Thus, in the final experiment, to assess whether male dominance interactions influenced female mate choice, in addition to infection, I tested female mate choice in a seminatural setting, which allowed for social interactions to occur. In this study, paternity was ascertained through DNA analysis and the resulting data indicated that neither male dominance status nor infection had a substantial impact on fema / Overall, the data do not support the theory that MHC-based preferences occur through urinary odours. However, the results do substantiate previous findings regarding female preferences for the odours of uninfected males. Furthermore, data from the controlled mate choice assay indicate that female odour preference for uninfected males extends to actual mate choice. Conversely, in the seminatural setting neither dominance nor infection prevailed as factors driving female mate choice. I suggest that this may be a consequence of the structural complexity of my arenas.

Bibliography : leaves 182-203. / 203 leaves : ill, map ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Describes the molecular analysis of the HLA class I and class II genes in the Australian Aboriginal population and also provides a comparison of serological and molecular tissue typing methods in view of genetic mutations at the HLA loci and the expression of serological HLA "blanks". / Thesis (Ph.D.)--University of Adelaide, Dept. of Medicine, 1996?

Thesis (Ph.D.] -- McMaster University, 1997. / Includes bibliographical references (leaves 142-155). Also available via World Wide Web.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 3 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 5 uppsatser.

Thesis (Ph. D.)--Northeastern University, 2008. / Title from title page (viewed March 3, 2009). Graduate School of Arts and Sciences, Dept. of Biology. Includes bibliographical references (p. 172-190).

Thesis (Ph. D.)--University of Nevada, Reno, 2008. / "May, 2008." Includes bibliographical references (leaves 99-110). Online version available on the World Wide Web.