A dedicated Molecular, Cell Biologist and Biochemist

Neeti P. Arora

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Home: 847-***-****, Mobile: 847-***-****

Email: *****.*****@*****.***

Summary of Qualifications

A hardworking and well-organized biologist/biochemist with broad research experience in industry and academics. I have independently executed complex molecular biology and bioanalytical projects involving mammalian and microbial systems.

• Cloning of the CDR1 promoter using luciferase (pCRW3), B-gal and GFP expression vectors, and subcloning using the pBS series of vectors for DNA sequencing and mutagenesis.

• Cloning of recombinant yeast and bovine F1 ATPase (using pRS3 series of shuttle vectors) and purification from 60 liters of yeast culture in a fermentor.

• Electrophoretic analysis, chromatography and Western analysis with crystallization of the F1 ATPase(structure solved at 2.8A)

• Primary cell culture of mesothelials from the parietal peritoneum of rats to study cytotoxicity of dialysis solutions by measurement of cytokines using ELISAs

• Worked with cell lines for human acute monocytic leukemia (THP1) and murine macrophage cell line (RAW) to establish them for use in cytotoxic studies.

• Currently working with PBMCs (Peripheral Blood Mononuclear cells) from humans and checking the levels of cytokines produced in response to exposure to Dialysis solutions.

• Initiated new projects at Baxter from concept to completion and published during PhD and Post-doc in peer reviewed journals-JBC, EMBO, FEMS, Acta Crystallographica etc.

• Data analysis using SigmaPlot (Standard Curve and Regression analysis)

• Authored Class B and Class C reports and drafted SOPs.

• Mentored PhD students in setting up the Protein purification lab at Chicago Medical School

Professional Experience

Research Scientist I

January 2009 – present, Renal Division, Baxter Healthcare through Kelly Services

• Study director of investigational and support protocols for Peritoneal dialysis.

• Independently plan, execute and manage projects that span multiple disciplines and utilize established methods, techniques or approaches.

• Work on the bench for isolation of human peripheral blood mononuclear cells (PBMCs) to study the release of biomarkers and check cytotoxicity of test solutions.

• Present data and share research outcomes with the group.

• Author Class B and Class C reports.

Research Associate

March 2007-Dec. 2008, Renal Division, Baxter Healthcare through Kelly Services

Solution Research and development for Peritoneal Dialysis

• Isolation and maintainence of rat primary mesothelial cells (first line of peritoneal host defense) as a model to check biocompatibility of new PD solutions

• Check response of mesothelial cells by production of biomarkers, viability, cytotoxicity

• Perform ELISAs from challenged cells for various biomarkers (MCP1, VEGF, TGF-beta, IL6, IL1, PAI 1, etc.)

Development of animal model for peritoneal transport (PET test)

• Handle rabbits and collection of dialysate via catheters at various time points

• Estimation of Intraperitoneal volume (IPV) using fluorescent volume marker for estimation of UF (Ultrafiltration)

• Perform various biochemical assays to determine the degradation of osmotic agent.

Research Associate

2002–2006 Department of Biochemistry and Molecular Biology, Rosalind Franklin University/Chicago Medical School, North Chicago

Protein purification, crystallization and reconstitution into liposomes.

• Developed the strategy for purifying multimeric enzyme, yeast mitochondrial ATP synthase, with optimal activity.

• Ascertained best conditions for crystallization of the protein (with all the 5 subunits) with a diffraction of 2.8Ao.

• Preparation of liposomes and reconstitution of proteoliposomes.

• Functional analysis of the reconstituted protein using various biochemical assays like ATPase.

Expression of bovine F1 ATPase with functional complementation in yeast Saccharomyces cerevisiae.

• Production of recombinant protein (ATP synthase) in yeast at fermenter scales.

• Purification of the protein using FPLC and affinity chromatography

Expression of Pseudomonas unc operon in E. coli to study its role in disease - cloning of genes using BAC DNA library

• PCR based site-directed mutagenesis - bacterial transformation, DNA isolation from BAC, bacteria and yeast, yeast transformation

Trained and mentored Ph.D students and other postdoctoral fellows on the protein purification and crystallization techniques.

PhD project

I was involved in studying the expression of a drug efflux pump, CDR1, of the pathogenic, multidrug resistant microbe Candida albicans. I identified regulatory regions in the upstream promoter region of CDR1 which play a key role in its overexpression in the presence of drug/stress.

Professional Skills

• Molecular Biology and Cell Biology: bacterial, yeast and mammalian (primary) cell culture, isolation of human peripheral blood mononuclear cells (PBMCs), maintenance and use of cell lines-THP1 and RAW, transformation, recombinant protein expression in prokaryotes (E.coli) and in eukaryotes (yeast), gene subcloning, DNA/RNA isolation and analysis, DNA modification and sequencing, primer design, PCR, northern, southern, site-directed mutagenesis, reporter assays (beta galactosidase and luciferase); gel shift assay, DNase I footprinting, Primer extension analysis

• Biochemistry: recombinant and native protein purification, SDS-PAGE electrophoresis, size exclusion and affinity chromatography, western blotting, protein quantification, bioanalytical assays for checking optimal enzyme activity, protein crystallization and freezing crystals, preparation of liposomes and reconstitution into proteoliposomes, drug testings (Disc and microtitre plate), ELISAs

• Clinical: Collection and processing of human blood and peritoneal effluent from rabbits and rats for chemical analysis, handling and checking well being of laboratory animals, injecting test solutions through an implanted access port and collecting samples at various time points, collecting tissue from animals

• Computer: SigmaPlot, MS office; sequence analysis and genome comparison (CLUSTALW, BLAST, NCBI GenBank and SWISS-PROT); EMBL, restriction map software (Webcutter), Transcription factor databases (MatInspector, TF search, TESS, TRANSFAC)

Education

Ph.D. 2001 Life Sciences, Jawaharlal Nehru University, New Delhi, India

M.S. 1996 Molecular Biology and Genetic Engineering, G.B. Pant University, India

Publications

Peer-reviewed journal publications

• Kabaleeswaran V., Puri N., Walker J.E., Leslie A.G.W. and Mueller D. M. Novel features of the rotary catalytic mechanism revealed in the structure of yeast F1 ATPase. EMBO J., 2006, 25(22), 5433-42

• Puri N., Lai-Zhang J., Mueller D. M. Expression of bovine F1 ATPase with functional complementation in yeast Saccharomyces cerevisiae. Journal of Biol. Chem. 2005, 280(23), 22418-24.

• Mueller D.M., Puri N., Kabaleeswaran V., Terry C., Leslie A.G., Walker J.E. Crystallization and preliminary crystallographic studies of the mitochondrial F1-ATPase from the yeast Saccharomyces cerevisiae. Acta Crystallogr D Biol. Crystallogr. 2004, 60(Pt 8): 1441-4

• Mueller D.M., Puri N., Kabaleeswaran V., Terry C., Leslie A.G., Walker J.E.

Ni-chelate affinity purification and crystallization of the yeast mitochondrial F1-ATPase. Protein Expression and Purification. 2004; 37(2): 479-85.

• Mueller, D.M., Puri, N., Kabaleeswaran, V., Terry, C., Leslie, A.G.W., and Walker, J.E. Structural analysis of the yeast F1-ATPase at a resolution of 2.8A. (2004) Advance Photon Source, Argonne National Laboratories, Argonne, IL

• Karnani N., Gaur N.A., Puri N., Krishnamurthy S, Goswami S.K., Mukhopadhyay G. and Prasad R. SRE1 and SRE2 are two specific steroid-responsive modules of Candida drug resistance gene 1 (CDR1) promoter. Yeast. 2004 Feb; 21(3): 219-39

• Gaur N. A., Puri N., Karnani N., Mukhopadhyay G., Goswami S. K. and Prasad R. Identification of a negative regulatory element, which regulates basal transcription of a multidrug resistance gene CDR1 of Candida albicans. FEMS Yeast Res. 2004, 4(4-5): 389-99.

• Puri, N., Krishnamurthy, S., Habib, S., Hasnain, S.E., Goswami, S. K. and Prasad, R. (1999) CDR1, a multidrug resistance gene from Candida albicans contains multiple regulatory domains in its promoter and the distal AP-1 mediates its induction by miconazole. FEMS Microbiol. Lett.180, 213-219.

• Gupta, V., Kohli, A., Krishnamurthy, S., Puri, N., Aalamgeer, S.A., Panwar, S.L. and Prasad, R. (1998) Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance and its in vitro transcriptional activation. Current Genetics, 34, 192-199.

Book Chapters

• Prasad, R., Panwar, S.L., Puri, N. (2001) Sex and Drug ATPases in fungi. In Pardha Saradhi, P. (Ed.) Biophysical Processes in Living Systems, Oxford and IBH Publishing Co. Pvt. Ltd.

• Singh, N.K., Sailesh, G., Kumar, A., Sarita, Puri, N., Singh, D.T., Garg, G.K., Nayal, S., Gaur,A.K., Srinivas,B.(1998) Improvement in the nutritional quality of wheat through genetic engineering. In Gupta, P.K., Singh, S.P., Balyan,H. S., Sharma, P.C., Ramesh, B. (Eds) Genetics and Biotechnology in Crop Improvement, Rastogi Publications Meerut, pp158-167.

Awards & Affiliations

• Poster titled- A rabbit model for the study of Icodextrin based peritoneal dialysis solution presented at ASN in October 2007.

• Travel award from Commonwealth and European Science Federation to present a poster entitled “Regulation of CDR1, an ABC drug transporter of Candida albicans” in the conference ABC 2001, “ABC proteins: From multidrug resistance to genetic disease" held in Gosau, Austria, 2001.

• Travel award from European Science Federation and the Department of Science and Technology, India to present a poster entitled “Multiple factors regulate the expression of CDR1, a multi drug transporter of C. albicans “ in the conference Human Fungal pathogens: Dimorphism and Disease held in Spain, 1999.

• Best poster award at the 67th Society of Biological Chemists meeting held at JNU, New Delhi.

• Best poster award in the “National Symposium on Plant Tissue Culture.” held in Pantnagar, India.

• CSIR/UGC National Educational Test 1995, Junior and Senior Research fellowships by Ministry of Human Resource and Development, Govt. of India (1996-2001).

• Graduate Aptitude Test Examination 1996, 93.6 percentile.

• Department of Biotechnology (DBT) fellowship, 1994-1996.

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