Defense Date

7-26-2004

Graduation Date

Summer 2004

Availability

Immediate Access

Submission Type

thesis

Degree Name

MS

Department

Biological Sciences

Committee Chair

John F. Stolz

Committee Member

Jana Patton-Vogt

Committee Member

Partha Basu

Keywords

arsenate reductase, arsenic, biochemical probes, microbial ecology

Abstract

To date, eighteen phylogenetically diverse prokaryotes have been found to utilize arsenic as the terminal electron acceptor in anaerobic respiration, converting arsenate into the more toxic and mobile arsenite. This process can lead to ground water contamination, making it imperative to detect in situ, active arsenate-respiring prokaryotes.

A biochemical probe that targets the catalytic subunit (ArrA) of the respiratory arsenate reductase (Arr) from Bacillus selenitireducens strain MLS10 was developed. Polyclonal antibodies were raised against a fifteen amino acids long highly conserved sequence at the C-terminus of ArrA. Highly specific antibodies obtained by affinity purification using the synthesized ArrA polypeptide, reacted with ArrA from B. selenitireducens, B. arsenicoselenatis, Clostridium sp. strain OhILAs, and strain SLAS-1, but not Bacillus sp. strain JMM-4, strain MLMS-1, and the epsilon Proteobacteria Sulfurospirillum barnesii, S.deleyianum, or S. arsenophilum. Western blot analysis and activity assays indicated that Arr from B. selenitireducens is up regulated by arsenic.

Format

PDF

Language

English

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