My research goal was to functionally dissect the regulatory region of the gene encoding the muscle protein troponin I (TnI) in the simple chordate Ciona intestinalis. Based on evolutionary sequence conservation and computational prediction of cis-elements in the regulatory region, I designed mutants and assayed their effects in a recombinant DNA construct reporter system. I electroporated TnI DNA-linked reporter constructs into Ciona zygotes and assayed reporter activity following 12-14 hr of development. These studies showed that a conserved E-box is essential for high-level embryonic tail muscle-specific expression whereas conserved MEF-2 and overlapping CREB/TEF-1 sites were not. I carried out functional promoter identification experiments and 5' RACE studies and localized the core-promoter/transcription start-site in the Ciona TnI gene. My results point out the similarities/differences between Ciona and vertebrate Tnl gene regulation, which presumably reflect the conservation/modification of TnI gene regulatory mechanisms during chordate/vertebrate evolution.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.84044 |
| Date | January 2005 |
| Creators | Khare, Parul |
| 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 (Department of Biology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 002269034, proquestno: AAIMR22736, Theses scanned by UMI/ProQuest. |
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