The creation and characterization of new plasmids by inserting specific DNA sequences of Ribonucleotide Reductases from bacteria into pMCSG7 and pMCSG28.

Arjun Deven , Annika Holmstrom, Youssef Sadek, and Shourya Sangam

Authors:  Youssef Sadek*, Arjun Deven*, Shourya Sangram*, Annika Holmstrom*

Faculty Mentor: Dr. Michael Harris

College:  College of Liberal Arts and Sciences 

Abstract

The activity of Ribonucleotide reductase (RNR) is regulated allosterically. This enzyme mediates the formation of deoxyribonucleotides in DNA synthesis by cleaving the 2’ -OH on ribonucleotides. To better characterize this regulation, we will create the plasmids that allow us to massively produce and purify the RNR protein. RNR genes from thermostable Thermatoga Maritima and pathogenic Pseudomonas aeruginosa were inserted into pMCSG7 and pMCSG28 vectors using Ligation-Independent Cloning. Then, the RNR enzyme will be expressed and purified. Finally, the activity and stability of the RNR enzyme will be characterized. Since the RNR is essential for the life cycle, the synthesized plasmids can serve to the studies of cancer and other pathogenic diseases.

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Alex Angerhofer
Alex Angerhofer (@guest_984)
1 year ago

Nice job documenting what you have done so far this semester.

Youssef Sadek
Youssef Sadek (@guest_3478)
Reply to  Alex Angerhofer
1 year ago

Thank you Dr. A.

Dr. Donnelly
Dr. Donnelly (@guest_1740)
1 year ago

Hopefully helpful suggestions: My job at the symposium is to ask students “why should I care” Very important with a general audience to make that clear up front. All figures should be numbered and labeled. Maybe that is a plasmid map, but what does that tell me? Would be good at a professional conference, nice lay out.

Youssef Sadek
Youssef Sadek (@guest_3596)
Reply to  Dr. Donnelly
1 year ago

Thank you Dr. Donnelly. We will definitely take your notes into consideration.

Arjun Deven
Arjun Deven (@guest_3788)
1 year ago

Thank you for the helpful suggestions! We will definitely update it with these in mind.