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October 03, 2013

Afia Dasgupta

******@*****.***, 806-***-****

EDUCATION

Center for Biotechnology and Genomics, Texas Tech University, Lubbock, TX, USA GPA: 3.84/4

MS in Biotechnology

RESEARCH INTERESTS

1. Cloning & subcloning - Restriction site analysis, Site-directed mutagenesis PCR, Ligation, Transformation,

Colony PCR, Plasmid DNA isolation

2. Protein Biochemistry - Transformation, over-expression of proteins, protein purification (nickel column, gel

filtation,ion-exchange & FPLC), SDS PAGE preparation and usage, Western Blot, Protein

quantification, Enzyme Assays and kinetics

3. Protein Secondary - Isothermal Titration Calorimetry, Circular Dichroism & Spectral perturbation

structure analysis

WORK EXPERIENCE

Graduate Research Assistant

Dr Knaff’s Research group, Center of Biotechnology and Genomics & Department of Chemistry and Biochemistry

Texas Tech University

“Illustrating the role of loop in Ferredoxin dependant Glutamate Synthase(Fd-GOGAT)”

Fd-dependant GOGAT was detected in chloroplasts of higher plants, cyanobacteria and green algae. The enzyme works by

binding to reduced Ferredoxin which delivers electrons via the 3Fe-4S cluster to FMN group. Fd-dependant Glutamate

synthases catalyze conversion of 2-oxoglutarate and glutamine to glutamate. The loop is present in plant type Ferredoxin

dependant GOGAT and is absent in NADPH dependant GOGAT. Preparation of both Wild type GOGAT enzyme and a loopless

variant of GOGAT from the cyanobacterium Synechocystis species PCC 6803

Contribution

- Performed overexpression and purification of the protein and comparison of characteristic properties such as km values for

glutamine, 2-oxoglutarate and Ferredoxin

- Obtained comparisons of prosthetic group content such as iron-sulfur and FMN content and the spectral properties.

- Conducted circular dichroism of the enzymes

- Used Isothermal Titration Calorimetry(ITC) to study complex formation and binding stoichiometry for both the enzymes

ACADEMIC PROJECTS

- Cloning, subcloning and overexpression of 3-alpha-OH-STEROID DEHYDROGENASE(CARBONYL REDUCTASE)

- Structural and functional analysis of Glutaredoxin after site-directed mutagenesis of Glutamate-34 to Leucine

INDUSTRIAL TRAINING

1. Aurigene Discovery Technologies Limited, Bangalore

2. Microbiology and Serology Lab, AMRI Hospital, Kolkata

MISCELLANEOUS

Best Speaker in “Bacillus Thurengiensis Brinjals are safe for India”.

(Reference available upon request)

(Geographically flexible)