Cloning a gamete fusion gene in Chlamydomonas reinhardtii.
Item
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Title
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Cloning a gamete fusion gene in Chlamydomonas reinhardtii.
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Identifier
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AAI3310666
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identifier
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3310666
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Creator
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Aksoy, Munevver.
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Contributor
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Adviser: Charlene L. Forest
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Date
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2008
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Molecular | Biology, Cell
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Abstract
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Many biological processes require cell fusion, therefore defects in cell fusion result in many diseases. The unicellular green alga Chlamydomonas reinhardtii is an excellent model organism to study cell-cell fusion. Objective of this study was to identify genes that are involved in Chlamydomonas mating type minus gamete fusion. A forward genetics approach was taken in our work. We created several mating type minus fusion defective mutants using DNA insertional mutagenesis. These mutants were normal in early stages of mating; they agglutinated with mating type plus, removed their walls, adhered to their mating partner through mating structure, but cells did not fuse indicating that the DNA insertional mutants were defective in the latest stages of fusion. The number of insertions was confirmed by Southern blots. Mutant J1 had one insertion and the flanking genomic DNA was cloned by TAIL-PCR and RESDA-PCR. We complemented the mutant J1 with a BAC that contained two genes; we were able to get zygotes when mated with wild type mating type plus cells. We proved that the progeny of these zygotes were products of meiosis. The BAC used for complementation had 2 genes, we need to determine which gene complemented the phenotype. We also identified a new mutant, with a phenotype that has never been reported in the literature. It agglutinates with itself and forms pellicles that look like zygote pellicle.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
