FLP-mediated conditional loss of an essential gene to facilitate complementation assays Metadata
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- Main Title FLP-mediated conditional loss of an essential gene to facilitate complementation assays
Author: Ganesan, SavitaCreator Type: Personal
Chair: Ayre, Brian G.Contributor Type: PersonalContributor Info: Major Professor
Committee Member: Chapman, Kent D.Contributor Type: Personal
Committee Member: Padilla, PamelaContributor Type: Personal
Name: University of North TexasPlace of Publication: Denton, Texas
- Creation: 2007-12
- Digitized: 2008-02-12
- Content Description: Commonly, when it is desirable to replace an essential gene with an allelic series of mutated genes, or genes with altered expression patterns, the complementing constructs are introduced into heterozygous plants, followed by the selection of homozygous null segregants. To overcome this laborious and time-consuming step, the newly developed two-component system utilizes a site-specific recombinase to excise a wild-type copy of the gene of interest from transformed tissues. In the first component (the first vector), a wild-type version of the gene is placed between target sequences recognized by FLP recombinase from the yeast 2 μm plasmid. This construct is transformed into a plant heterozygous for a null mutation at the endogenous locus, and progeny plants carrying the excisable complementing gene and segregating homozygous knockout at the endogenous locus are selected. The second component (the second vector) carries the experimental gene along with the FLP gene. When this construct is introduced, FLP recombinase excises the complementing gene, leaving the experimental gene as the only functional copy. The FLP gene is driven by an egg apparatus specific enhancer (EASE) to ensure excision of the complementing cDNA in the egg cell and zygote following floral-dip transformation. The utility of this system is being tested using various experimental derivatives of the essential sucrose-proton symporter, AtSUC2, which is required for photoassimilate transport.
- Keyword: FLP/FRT recombination
- Keyword: complementation assay
- Keyword: ATSUC2
- Library of Congress Subject Headings: Gene targeting.
- Library of Congress Subject Headings: Genetic engineering.
- Library of Congress Subject Headings: Complementation (Genetics)
- Library of Congress Subject Headings: Genetic recombination.
Name: UNT Theses and DissertationsCode: UNTETD
Name: UNT LibrariesCode: UNT
- Rights Access: public
- Rights License: copyright
- Rights Holder: Ganesan, Savita
- Rights Statement: Copyright is held by the author, unless otherwise noted. All rights reserved.
- Thesis or Dissertation
- OCLC: 228010560
- Archival Resource Key: ark:/67531/metadc5180
- Degree Name: Master of Science
- Degree Level: Master's
- Degree Discipline: Biochemistry
- Academic Department: Department of Biological Sciences
- Degree Grantor: University of North Texas