Author

Lina Bird

Defense Date

6-13-2007

Graduation Date

2007

Availability

Immediate Access

Submission Type

thesis

Degree Name

MS

Department

Environmental Science and Management (ESM)

School

Bayer School of Natural and Environmental Sciences

Committee Chair

John Stoltz

Committee Member

Nancy Trun

Committee Member

Partha Basu

Keywords

molybdoenzyme, over expression

Abstract

Periplasmic nitrate reductase from Sulfurospirillum barnesii is representative of the distinct branch of epsilonproteobacteria periplasmic nitrate reductases. The gene encoding the active subunit of S. barnesii's periplasmic nitrate reductase (napA) was successfully cloned into the vector pET101/D/TOPO. Constructs were made both with and without the sequence coding for the periplasmic signal sequence. Preliminary expression and purification experiments were performed with limited success. The DNA encoding the periplasmic signal sequence proved necessary for expression. Even with the signal sequence coding region, protein expression was limited. The low levels of expression appeared due to product degradation during cell growth. Purification was incomplete, with both low recovery levels and significant contamination.

Format

PDF

Language

English

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