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

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.

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