Cloning and Overexpression of Yeast Cystathionine γ-Lyase

Cystathionine γ-lyase (CGL) is a protein involved in the conversion of methionine to cysteine as part of the trans-sulfuration pathway. Inactivation of the enzyme, possibly by oxidation of reactive site residues, has been linked to cataractogenesis. This research isolated the cystathionine γ-lyase gene (<i>CYS3</i>), modified it with an XhoI and BlpI restriction site and enterokinase sequence (creating a modified <i>CYS3</i>, <i>mCYS3</i>, gene), and ultimately spliced it into a pET-15b plasmid to create a recombinant plasmid. Then transformed the recombinant plasmid into BL21 E. coli cells. This enabled the production of bacterial cells capable of generating µg amounts of CGL for future metal catalyzed oxidation (MCO) research.

Name thesis:degree_name

Master of Science in Chemistry

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

Department of Chemistry

Grantor dc:publisher

Youngstown State University

Year dc:date

2012

Author dc:creator

• Raby, Roger Lee, Jr.

Contributors dc:contributor

• Serra, Michael

• Biochemistry
• Chemistry
• cloning
• overexpression
• cataractogenesis
• cystahionine gamma-lyase
• enterokinase
• plasmid
• cystathionine

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=ysu1355527551

OAI identifier oai:identifier

oai:etd.ohiolink.edu:ysu1355527551