dr. mueller’s mechanistic enzymology lab chemistry and biochemistry department

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Name: Ms. Marguerite McDonald Title: Biochemist, grad student Concentration: Enzymology Work Place : Dr. Mueller’s Mechanistic Enzymology Lab Chemistry and Biochemistry Department University of Delaware Graduate Program

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A Day in the Lab with. Name: Ms. Marguerite McDonald Title: Biochemist, grad student Concentration: Enzymology Work Place :. Dr. Mueller’s Mechanistic Enzymology Lab Chemistry and Biochemistry Department University of Delaware Graduate Program. PROTEIN. RNA. DNA. The Central Dogma. - PowerPoint PPT Presentation

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Page 1: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

Name: Ms. Marguerite McDonald Title: Biochemist, grad studentConcentration: EnzymologyWork Place :

Dr. Mueller’s

Mechanistic Enzymology Lab

Chemistry and Biochemistry Department

University of Delaware Graduate Program

Page 2: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

Biochemistry

DNA RNA PROTEIN

http://bioweb.wku.edu/courses/BIOL115/Wyatt/Nucleic_Acids/mRNA2.htm

Page 3: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

Mechanistic EnzymologyThe study of the action (the mechanism) of an enzyme

http://www.sdnhm.org/exhibits/epidemic/teachers/images/bacteria.jpg

Pseudouridine synthases

synthase

synthase

Page 4: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

The Chemistry: Isomerization of RNA

Page 5: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

TruA’s substrate: transfer RNA

Page 6: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

Purifying radioactive tRNA and TruA

urea-polyacrylamide gel showing pure tRNA and tRNA with DNA contamination

SDS-polyacrylamide gel showing pure TruA and TruA before purification

HPLC trace and radioactivity/ min

0

50000

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350000

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450000

0 5 10 15 20 25

time

dpm

HPLC tracedpms/10 ul fraction

Page 7: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department
Page 8: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department
Page 9: Dr. Mueller’s  Mechanistic Enzymology Lab Chemistry and Biochemistry Department

Pseudouridine Synthases