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RESUME

Personal Information:

Srinivas Rampalli

**** ***** ******** **** ****** : 773-***-****.

Apt.206, Chicago E-Mail: *********@*******.***

Illinois 60640.

Work Permit: US citizen.

Managerial experience: I have two technicians (One from cell culture area and one from cloning area) reporting to me. During my post-doctoral studies, two technicians were reporting to me (one from cell culture area and one from recombinant viral construction lab). I have helped and guided several interns in their completion of summer projects.

Work experience:

2007-Present: Scientist (Molecular biology) in R&D systems, Minneapolis

Job responsibilities: -

Challenging protein expression in CHO-S, HEK293, NSO, Per C6 cell lines. Bone morphogenetic protein expression using above cell lines. Semaphorin protein expression. Contribute to the development of new products by incorporating technologies related to recombinant protein expression in eukaryotic cells. My other responsibilities focus on analyzing the structure and characteristics of complex proteins in order to design efficient recombinant expression strategies. This may include gene modification as well as possible design and construction of expression vectors used for a broad range of protein expression in multiple systems. I also interact with other departments in order to facilitate effective progress of candidate reagents through various stages of product development.

Achievements: Solved the expression problems of Semaphorin 3 and 4 family members, Solved expression problems of hBMP-15, mGDF-5, m/h-GDF-3, hBMP-3. Solved expression problems of pro-convertases (PC1, PC2, PC4, NRD1, hMBTPS1,PCSK9)

2005-2006 Molecular biologist (contractor)/Protein purification scientist in protein

biochemistry department in ABBOTT labs, IL.

Job responsibilities:

Cloning, expression of (E.coli, Pichia vector construction and expression, insect cells, mammalian cells) and subsequent protein isolation of N-His and C-His proteins and characterization, Performing all standard molecular biology laboratory procedures including Gateway cloning, 96 well cloning procedures, synthetic gene construction and site directed mutagenesis. Day to day use of VectorNTI,, as well as expertise in protein isolation and characterization using standard biochemical laboratory techniques. Additional responsibilities include providing regular updates, documentation, and communication of project results as appropriate, participating in national scientific meetings, and keeping current with field of expertise, including literature and technology development in the area of molecular biology.

Achievements: Developed high throughput baculo viral vectors using 96-well transfection method. Expressed several proteins using baculo viral vectors. Expressed several proteins using pET vectors in E.coli.

2001-05 Senior Research Associate in CMRC

Feinberg Medical School, Chicago.

1997-01 Research Associate in Department of Microbiology and Immunology. Feinberg Medical School. Chicago.

1996-97 Post Doctoral Fellow with Dr.Hans Lehrach, Max Planck Institute fur Molekulare Genetik, Berlin, Germany.

1989-96 PhD in Biochemistry from University Hyderabad, India

1986-89 Worked as Microbiologist in Stangen Immunodiagnostics (Dr. Reddy group of

Companies, Hyderabad, India.

Advanced certificate in Regulatory affairs (date of completion June 2006):

RA 601 - Introduction to the Pharmaceutical, Biotechnology and Medical Device Industries (Completed).

RA 602 - Food and Drug Law (Completed).

RA 770 - Current Good Manufacturing Practices -General Concepts (Completed).

RA 774 – Investigational and marketing Applications for Drugs, Biologics and Medical devices (Planning to take in future).

QSR Training: Quality system regulation for medical devices.

Viral clearance: Procedures for Biopharmaceutical safety: Course deals with viral contamination of biologicals, controlling production processes for viral safety, Viral detection methods, managing risks to viral safety and process validation for viral clearance.

Professional Accomplishments:

· Constructed several adenoviral vectors for other distinguished scientists and transferred the material through material transfer agreements.

· Worked as mentor for two graduate students for their summer internship.

· Worked as a tutor for candidates aspiring for a degree in Medicine in India.

Fellowships and Awards

2003 MRIC-CMIER conference award for Gene therapy meeting.

1996 Post Doctoral Fellowship from Max Planck Institute fur Molekulare Genetik, Berlin, Germany.

1991 Senior Research Fellowship from University Grants Commission, India.

1989 Junior Research Fellowship from University Grants Commission, India.

1986 Awarded Gold Medal in Master’s Degree.

Bioinformatics Skills

Operating systems: Macintosh, Familiarity in working in UNIX and Windows environments.

Databases: Microsoft Access 97, Oracle 7.3 (Creation of databases, retrieval of information using SQL queries), Proficient in using Software’s like GCG and data retrieval systems at NCBI. Usage of other analytical programs like restriction map software (web cutter), Transcription factor databases (TF search, TESS, TRANSFAC) and Multiple sequence alignments (Clustal, Macaw), Vector NTI, Contig express and Align X from Vector NTI, DNA Strider.

Computer applications: Microsoft word, Excel, PowerPoint and Adobe Photoshop

Research Experience

Molecular Biology techniques:

Preparation of stable cell lines expressing sense and antisense strands of cDNA, Isolation of cosmid and genomic DNA, Electro elution of proteins and from gels, cloning of specific sequences into plasmids and bacteriophages, over expression and purification of ribosomal proteins in pET vectors. Site directed mutagenesis, Gel mobility shift assays, and Taqman quantitative PCR and Affymetrix micro array analysis. Yeast two- Hybrid assays and Immunoprecipitations for protein-protein interaction studies. Protein over expression systems using T7, pET, GST vectors in Bl21, XL1 and DH5 alpha cells and GST pull down assays, Shotgun cloning of genomic sequences for protein expression libraries. Preparation of cDNA, PCR, RT-PCR and QRT-PCR. Transient and stable transfections. TNT T7 coupled transcription and translation. Southern, northern and western blotting, DNA foot printing and colony hybridization. Isolation of mRNA from prokaryotic and eukaryotic cells. Transformation of yeast and over expression of proteins in yeast for protein-protein interaction studies,

Virology Techniques (Baculo virus, Adeno virus and Adeno associated virus):

Adeno viral construction using homologous recombination and ligation methods, Viral purification using CSCl centrifugation, Plaque assays and recombinant vector detection. Construction of several adeno associated viral vectors, viral purification (Iodoxinol step gradient centrifugation and FPLC method) and titering using ELISA and QRT-PCR. In vitro reporter assays and regulation (flow cytometry and QRT-PCR for detecting mRNA copy number) using regulatable promoters. Construction of baculoviral vectors for protein expression and AAV production. Growth of different types of bacteriophages, purification of DNA from different types of bacteriophages, cloning prokaryotic DNA into bacteriphage arms and packaging to form library, titering and plating of packaged library and library amplification. Growth and maintenance of ATCC 293, U20S, Hela, MCF10A, CHO, COS, 3T3 and SF9 cell lines. Over expression of proteins using adeno viral and baculo viral vectors. Construction of recombinant baculovirus vectors for making AAV vectors. Over expression of proteins in mammalian cells using CMV, SV 40 promoters, Infection of neuronal cell lines using replication defective lent viral vectors.

Protein chemistry techniques:

Isolation of ribosome’s from different bacteria (eubacteria, archaebacteria and eukaryotes). Isolation of ribosomal subunits using gradient centrifugation (vertical and horizontal), 2D- gel electrophoresis, IEF, NEPHGE, Geyl et al., Madjar et al., and Mets and Bogorad methods. in vitro reconstitution of ribosome’s and in vitro translation. HPLC purification of proteins. Quantitation of proteins using ELISA. Fractionation of ribosomal proteins using antibiotic novobiocin and hydrophobic chromatography. Spectroscopy and Thermal melting analysis of DNA and proteins. Protein folding studies using fluorescence spectrophotometer, Cell cycle analysis and apoptosis using flow cytometry.

Biochemical Techniques:

Protein purification using Ultracentrifugation, Salt Precipitation, Chromatography(affinity, Ion exchange and gel filtration), Ultrafiltration of ribosomal proteins, Cyanogen bromide activation of sepharose and covalent coupling, Electrophoresis of proteins under native and denaturing conditions and on gradient gels. Preparation of monospecific polyclonal antibodies, purification of IgG through chromatography on protein A sepharose, Western blotting using HRP and Apase systems. Radiometric Techniques: Scintillation counting and autoradiography.

Animal surgeries: In vivo skills include handling of animals, behavioral testing and stereotaxic surgery methods. Injection of AAV vectors in to striatum.

Publications:

1. R Srinivas, K Sreenivas and T Suryanarayana (1992) Novel fractionation of proteins using novobiocin as a precipitating agent: Application of ribosomal proteins from the archaebacterium Sulfolobus acidocaldarius and E.Coli. Biochem.International.Vol.27:221-229

2. KSG Haviryagi, R Srinivas, T Suryanarayana and Mohan C Vemuri (1992) Effects of ethanol on hepatic ribosomal proteins and mRNA. Molecular and Cellular Biochemistry.Vol.115:143-147.

3. G. Jayaraman, R. Srinivas, C. Duggan, E. Ferreira, S. Swaminathan, K. Somasundaram, J Williams, C. Hauser, M. Kurkinen, R. Dhar, S. Weitzman, G. Buttice and B. Thimmapaya. (1999). (Both first and second authors contributed equally towards the work)

p300/cAMP responsive element-binding protein interactions with Ets-1 and Ets-2 in the transcriptional activation of the human stromelysin promoter. Journal of biological chemistry. Vol.274:173**-*****.

4. Karthikeyan Narayanan, Rampalli Srinivas, Amsaveni Ramachandran, Jianjun Hao, Bruce Quinn and Anne George.

Differentiation of Embryonic Mesenchymal cells to Odontoblast-like cells by over expression of Dentin matrix protein 1. 2001, Proceedings of Natural Academy of Sciences (PNAS, USA) (2001) Vol: 98: 4516-4521.

5. Fang M, Rampalli S, Suddoth J, Sweet A, Gottardi-Little F, Quinn B. Novel adenovirus constructs for dementia-related proteins: Tau and APP adenovirus vectors. Journal of the American Geriatrics Society. Vol. 49(4): P209 Apr 2001.

6. Effects of Depletion of CBP on c-Myc Regulation and Cell Cycle G1-S

Transition. Published on November 2, 2004 as Manuscript M408633200

Hasan N. Rajabi, Sudhakar Baluchamy, Sivanagarani Kolli , Alo Nag, Rampalli

Srinivas , Pradip Raychaudhuri and Bayar Thimmapaya

7. Lixin Jiang, Srinivas Rampalli, David George, Craig Press, Eric G. Bremer,

Maurice R.G. O'Gorman, Martha C. Bohn.

Tight Regulation from a Single rAAV Vector as Demonstrated by Flow Cytometry

and Quantitative RT-PCR. Gene therapy(2004). Vol 11, 1057-1067.

8. Sodhi, C., Rampalli Srinivas, Perez Ruth, Koo ED, Allison Sweet, Bruce Quinn, Gottardi-Littell

The endocytotic pathway is required for increased Abeta42 secretion during apoptosis. Molecular brain research 128 (2004) 201–211.

9. Karthikeyan Narayanan*, Rampalli Srinivas*, Mathew Craig Peterson, Amsaveni Ramachandran, Jianjun Hao, Bayar Thimmapaya, Philipp E. Scherer and Anne George (Both first and second authors contributed equally towards the work).

Transcriptional regulation of Dentin Matrix Protein 1 by JunB and P300 during osteoblast differentiation.

Accepted for publication JBC August 11, 2004.

Abstracts and Posters:

1.Presented an abstract in Seventeenth European Crystallographic meeting held at Instituto Superio Tecino in Lisbon, Portugal. Ribosomal Crystallography: Advances in Phasing. 24-8-97 to 28-8-97

2. Presented an abstract in 4th Triannual meeting on ribosome synthesis

The Ribosome: It’s(Nucleolar) synthesis and structure.16-8-97 to 20-8-97, Leiden Institute of Chemistry, leiden University, Gorlaeus laboratories, Netherlands.

3. Presented an abstract in RNA structure: CMB/RNA and RNA society, Sinsheimer Labs, University of California, California, June 25-29, 1997.

4. Presented an abstract at Chicago, USA. For American Geriatrics Society: Novel Adenovirus constructs for dementia related proteins: Tau and APP adenovirus vectors.

5. Presented an abstract and poster at Boston, USA. For American society for Gene Therapy (2002).

“Quantitation of transgene expression from self-regulating rAAV vectors by flow cytometry”.

L. Jiang, S. Rampalli, M.R.G. O’Gorman, M.C.Bohn.

6. Presented an abstract and poster to Society of Neuroscience annual meeting at Orlando, Florida from Nov-2-7, 2002.

Tight regulation from a rAAV vector as demonstrated by flow cytometry and Quantitative RT-PCR.

L. Jiang; S. Rampalli; D. George; C. Press; E.G. Bremer; M.R.G. O'Gorman; M.C. Bohn.

7. Presented an abstract and poster to Society of Neuroscience annual meeting at Orlando, Florida from Nov-2-7, 2002.

Quantitative assays of humanized green fluorescent protein (hGFP) expression from recombinant “tetracycline (tet) on” adeno-associated vectors (rAAV).

S. Rampalli, L. Jiang, C. Press & M.C. Bohn.

8. Presented an abstract to Society of Neuroscience annual meeting at New Orleans, Louisiana from Nov 8 -12, 2003.

Tight regulation of AADC and GFP expression from AAV vectors harboring a Tetracycline (TET)-on regulated promoter.

S. Rampalli, L. Jiang, S. Barlow & M.C. Bohn.

9. Submitted an abstract to Society of Neuroscience annual meeting at New Orleans, Louisiana from Nov 8 -12, 2003.

Tight regulation from rAAV vectors as demonstrated by flow cytometry and quantitative RT-PCR both in vitro and in vivo

L. Jiang., S. Rampalli., D. George., E.G. Bremer., M.R.G. O'Gorman and M.C. Bohn.

10. Presented an abstract and poster at Washington D.C, USA. For American society for Gene Therapy (2003).

Tight regulation from rAAV vectors as demonstrated by flow cytometry and quantitative RT-PCR both in vitro and in vivo

L. Jiang, S. Rampalli, C. Press, D. George, E. Bremer, M.R.G. O’Gorman, M.C.Bohn.

11. Presented an abstract and poster at Washington D.C, USA. For American society for Gene Therapy (2003).

rAAV-2 vectors containing tet-on elements display tight regulation as assessed by flow cytometry and real-time RT-PCR.

S. Rampalli, L. Jiang, S. Barlow & M.C. Bohn,

12. Submitted an abstract and poster at Society of Neuroscience annual meeting at New Orleans, Louisiana from Nov 8 -12, 2003.

The endocytotic pathway is required for increased abeta42 secretion during apoptosis.

C.P. Sodhi, S. Rampalli, R.G. Perez, E.H.Koo, B. Quinn and N.R. Gottardi-littell.

13. Poster presentation at Society of Neuroscience annual meeting in San Diego, California from Oct 23-27, 2003.

Expression of aromatic amino acid dopa decarboxylase (AADC) from a self-regulatable rAAV2 vector is effectively regulated in rat brain.

S. Rampalli, N.C. West, L. Jiang, and M.C. Bohn.

14. Poster presentation at Children’s Memorial Research Center, Chicago. May 26 2005.

Gene regulation of aromatic amino acid dopa decarboxylase (AADC) from Tet-on and Tet-off self-regulatable rAAV2 vectors.

S. Rampalli, N.C. West, L. Jiang, and M.C. Bohn.

15. Poster presentation at Children’s Memorial Research Center, Chicago. May 26 2005.

AAV delivery of human GDNF for treatment of Parkinson’s disease.

N.C. West, S. Rampalli, L. Jiang, and M.C. Bohn.

Work shops and meetings:

1. Nucleic Acid and protein sequence analysis work shop conducted in Feinberg Medical School on February 1999.

2. A field guide to Genebank and NCBI resources, work shop conducted in Northwestern Medical School on April 28,1999.

3. Genome sequencing and Biology workshop conducted in ColdSpring harbor laboratory on May 19-23, 1999.

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