Zinc cluster proteins represent a subclass of zinc finger proteins and function as transcriptional regulators. An in vivo genetic screening system was developed in yeast to identify DNA binding sites and specificities for these proteins. / An oligonucleotide library of 200 000 clones was constructed. Control screening trials with Hap1p and Gal4p demonstrated effectiveness in recovering binding sites. Sequencing of isolated clones showed correlation with published target sequences and binding was confirmed by electrophoretic mobility shift assay (EMSA). / Screening over 100 000 clones of the library with the YLR228c gene product allowed the isolation of 10 clones. Mutational EMSA studies were performed to identify nucleotides important for binding to derive a consensus sequence. A CGG triplet was found to be significant for binding. It can be hypothesized that Ylr228p may bind as a monomer to its targets.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.30813 |
| Date | January 2000 |
| Creators | Chow, Christine, 1974- |
| Contributors | Turcotte, Bernard (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 | Master of Science (Division of Experimental Medicine.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001804648, proquestno: MQ70402, Theses scanned by UMI/ProQuest. |
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