Title

Purification and preliminary characterization of Zinc(II) dependent transcription factor AdcR

Author

ASM Saem

Advisor

Emerson, Joseph P.

Committee Member

Fitzkee, Nicholas C.

Committee Member

Mlsna, Debra Ann

Date of Degree

8-1-2019

Original embargo terms

Visible MSU Only 3 Years||8/15/2022

Document Type

Graduate Thesis - Open Access

Major

Chemistry

Degree Name

Master of Science

College

College of Arts and Sciences

Department

Department of Chemistry

Abstract

Transition metals are essential micronutrients for organisms. Excess zinc can be lethal to bacterial survival. Zinc homeostasis is crucial and is regulated by zinc uptake and efflux system. Streptococcus pneumoniae AdcR (Adhesin competence regulator) transcription factor is the member of MarR (multiple antibiotic resistance regulator) family proteins that suppress the zinc upregulation transporter. In the absence of zinc, AdcR has less affinity to bind DNA and uptake genes are expressed in order to transport zinc inside the cell. Streptococcus pneumoniae is a gram-positive human pathogen that is capable of colonizing in the human nasopharynx, especially in young children and leads to severe diseases including pneumonia, septicaemia, and meningitis. In this work, we have transformed AdcR gene in E.Coli (BL21D3) cell, and we have overexpressed and purified AdcR using classical molecular biology and protein purification techniques. We also performed preliminary biophysical characterization of purified zinc(II) dependent transcription factor AdcR.

URI

https://hdl.handle.net/11668/14572

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