Date of Award

1998

Degree Type

Thesis

Degree Name

MS Biology

Department

Biology

Advisor

Carolyn Bentivegna

Keywords

Biology, Balbiani ring gene, Molecular probe

Abstract

The Environmental Protection Agency is currently developing sediment quality criteria in order to protect the nation's water sources. In order to assist in the development of standards for the disposal of heavy metals, our laboratory is undergoing sediment toxicity studies on Chironomus riparius. It is an EPA approved test organism that spends most of its life in the sediment. Following a previous study (Cochrane et al., 1994), the organism's response to heavy metals will be monitored on the genetic level. Several genes, including a Balbiani ring (BR) gene, which was the focus of this thesis, will be studied to determine the effects on carbohydrate metabolism and induction of protective proteins. Much of the genome of C. riparius is unknown, therefore, the genes involved in toxicity studies had to first be sequenced. Primers to amplify a region of a BR gene were selected from a species within the same Genus, C. tentans. Sequencing of the amplified fragment was based on the Sanger sequencing method. The experimental sequence was compared to a database for confirmation. Molecular probes based on the experimental sequence were made and then tested for specificity and ability to detect varying levels of message using solution hybridization and differential display. A Balbiani ring fragment was isolated and sequenced, but the probe was unable to detect message. It is possible that the message was not induced.

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