X-Linked Gene Expression in Wild-Type, rha-1(tm329), and rde-2(ne221) C. elegans Using Realt-Time Reverse Transcription Polymerase Chain Reaction

Author

Justin White

Date of Award

2007

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Keywords

C. elegans, RHA-1, Real-Time RT PCR

Area of Concentration

Biochemistry

Abstract

Helicases are enzymes that separate dsRNA or dsDNA and have been implicated in several different cellular processes. In particular, RNA helicase A (RHA) is a multifunctional helicase involved in transcription regulation and histone modification. A deletion in rha-1 in C. elegans produces a temperature sensitive sterile mutant (Walstrom et al., 2005). The mutant worms display a different series of histone modification and of particular interest is the loss of the histone 3 dimethylation on lysine 9. Rha- 1 also plays a role in the conserved RNA interference (RNAi) pathway, which is a post-transcriptional silencing pathway. Another gene required for RNAi is rde-2. We used a null mutant of rde-2 and the rha-1 mutant to determine the respective effects of the mutations on genetic expression of X-linked genes in the germline. Using Real-time RT-PCR, we discovered that there was little change the expression of many of the X-linked genes examined. There was more expression in the rha-1 mutants than in the rde- 2 mutants, which can be explained by the presence of the H3K9 methylation (silencing modification) in the rde-2 mutants but not in the rha-1 mutants. There seemed to be a defect in silencing with respect to gene T08D2.4 in both mutant strains. After examining the gene expression results, a double mutant was constructed.

Rights

This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

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