Identification and Characterization of A Zebrafish Glutathione S-Transferase Pi

Glutathione S-transferases (GSTs) are a major group of Phase II enzymes involved in the detoxification of certain endogenous compounds such as reactive oxygen species (ROS) generated during metabolism, as well as in the detoxification of xenobiotics including drugs. In recent years, the zebrafish has increasingly been used as a promising animal model for biomedical research. This study is part of an effort to establish the zebrafish as a model for studying GST-mediated glutathione conjugation of xenobiotics. By searching the GenBank database, we have identified sequences encoding fifteen zebrafish GSTs. Using a computer algorithm available at the Genebee website, a dendrogram of the fifteen zebrafish GSTs was generated. A zebrafish GST Pi was subsequently cloned, expressed, and purified. An enzymatic assay for purified GST Pi was established using 1-chloro-2, 4-dinitrobenzene (CDNB) as a substrate. A pH-dependency study was performed, and the results indicated that the zebrafish GST Pi was active over a pH range of 6 and 8 with optimal activity observed at pH 7.5. The expression of novel zebrafishGST Pi during various developmental stages from embryogenesis to maturity was examined via reverse transcription-polymerase chain reaction (RT-PCR).

Name thesis:degree_name

Master of Science in Pharmaceutical Science (MSP)

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

College of Pharmacy

Grantor dc:publisher

University of Toledo Health Science Campus

Year dc:date

2013

Author dc:creator

• Abunnaja, Maryam

Contributors dc:contributor

• Liu, Ming-Cheh

• Pharmacology

Statement dc:rights

• unrestricted
• This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.

Language dc:language

English

Repository record dc:identifier

http://rave.ohiolink.edu/etdc/view?acc_num=mco1364857296

OAI identifier oai:identifier

oai:etd.ohiolink.edu:mco1364857296