As part of a study investigating fundamental cellular immunology in the sheep, this thesis describes the characterisation and expression of the genes of the sheep MHC class II region. Cosmid libraries prepared from DNA from three unrelated sheep were screened with probes from the <I>DP, DQ</I> and <I>DR</I> sub-regions of the human and mouse MHC class II regions. Cosmids were used because they facilitate the cloning of relatively large genomic inserts. Restriction maps of the cosmids have been produced showing that some of the clones overlapped. The MHC <I>A</I> and <I>B</I> genes within the clones have been sequenced and assigned to a specific sub-type. Functional genes have been identified by the reaction of their products with anti-sheep class II monoclonal antibodies following DNA-mediated transfection into the mouse L cell, a fibroblast cell line which does not express endogenous mouse class II genes. Transcription of some of the genes has been demonstrated by Northern blots and reverse transcription polymerase chain reaction. A restriction map of the sheep class II <I>DQ</I> sub-region has been constructed and shown to contain two distinct <I>DQA</I> loci with associated <I>DQB</I> genes. The <I>DQ1</I> <I>A/B</I> gene pair was expressed in the mouse L cell. The sheep class II <I>DQ1</I> product at the cell surface reacted with a sub-set of the available anti-sheep class II monoclonal antibodies. The <I>DQ2</I> genes were transcribed and some evidence for their cell surface expression was obtained, although this was not formally proved.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:664034 |
| Date | January 1996 |
| Creators | Wright, Harry |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/30049 |
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