Return to search

Construction and application of specific microsatellite-enriched DNA library

To facilitate the isolation and characterisation of polymorphic markers for the Poultry Genome Mapping project, and to define potential transcriptional factors or disease candidate genes, the following studies were carried out as the purpose of this thesis. Procedures were developed for the construction of specific microsatellite-enriched DNA libraries. [CA/TG]<SUB>n</SUB> microsatellite-enriched genomic DNA libraries were constructed through marker selection and magnetic particle selection respectively; whereas magnetic particle selection was used in the construction of [CA/TG]<SUB>n</SUB> and [CAG/CTG]<SUB>n </SUB>microsatellite enriched liver cDNA libraries. Positive clones in the DNA libraries constructed ranged from 0.5% to 5% depending on the type of microsatellite repeat. Two hundred microsatellite positive clones from genomic DNA library and a hundred from liver cDNA library were identified by screening once with an oligo probe and further characterised by sequencing. Polymorphisms of these microsatellites were studied by polymerase chain reaction and mapped using the EAST LANSING and COMPTON mapping cross. A nonredundant database search using the available expressed sequence information revealed chicken homologous sequences of human transcriptional factor gene, MEF2 and human Fragile X gene, FMR1 as well as a substantial number of new genes. Chromosome mapping and study of differential expression of these genes are underway. The isolation of these microsatellites will facilitate the mapping of important chicken quantitative traits, e.g. production traits. The mapping of expressed sequences will help to define candidate genes of these and other traits.

Identiferoai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:654014
Date January 1994
CreatorsLong, Q.
PublisherUniversity of Edinburgh
Source SetsEthos UK
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation

Page generated in 0.0024 seconds