RESUME – Scientist/Professor in Molecular Biology

Premraj Pushpakaran, PhD

Residence: 36/86b, Pranavam, Manthra Parambil H, Kotte Canal Road, Kochi - 682 018,Kerala (India) Ph: (91) 484-2403539; e-mail:ppushpakaran@gmail.com,premraj74@rediffmail.com---------------------------------------------------------------------------------------------------------------------Career Objective: Seeking a research oriented position that utilizes my expertise in Proteinchemistry, molecular and diagnostic microbiology, and eukaryotic gene expression andregulation to grow in a mutually beneficial manner.Professional Summary: 10 years molecular biology, over 10 years microbiology, 4 yearsproject management: Cloning, expression and purification of proteins, development of molecular diagnostics, microarray analysis and data mining in research labs. Specialtiesinclude: multi-disciplinary project leadership, strong knowledge of the microbiology,molecular biology, clinical analyses, scientific software, and protein chemistry.

Skills:Project Management

Grants support, Research design, scientific collaborations.Group Leader, Managed a team of technicians and undergraduate students.

Scientific Expertise

Molecular Techniques

: Microarray analysis, DNA, RNA handling, Cloning, PCR, RT-PCR (Real-time and agarose), Southern, cytotoxicity assays, Northern analysis (radioactive and non-radioactive chemistries).

Genotyping

: PCR and restriction analysis based genotyping for

Mycobacterium tuberculosis

,HBV, HCV, RSV. RFLP and RAPD typing of Juniper plant sp.

Protein Expression and analysis

: Recombinant proteins (GST, His Tag) expressed in bacterialand baculovirus/Sf-9 cells/High-Five cells. Column purified several proteins includingglycoproteins (soluble forms or from pellet).

Signal transduction pathways

: deciphering using Kinase assays, Immunoprecipitation, Westerns.

Antibody development

: polyclonal and monoclonal antibodies

Immunoassay development

: ELISA for serum IgG and tear secretory IgA detection.

Microbiology

: Isolation, identification and typing of viruses, fungi and bacteria.

Bioinformatics/Application Software

DNA Star, Vector NTI, Oligo 4.0, Primer 3.0, Fast PCR, Affymetrix Microarray Suite 5.0, DataMining tool, GeneSpring, GenMAPP, Sigmaplot, SEQUSET, MASCOT, Adobe PhotoShop, MSExcel, PowerPoint, Word, Office 2003.

Entrepreneurial Skill

Clinical microbiology laboratory

: Established a Diagnostic microbiology Lab for 200-beddedhospital with capabilities such as bacterial isolation, identification, sensitivity testing (including

M. tuberculosis, H. pylori

), blood bank screening for HBV, HCV, HIV, VDRL. Abbott ELISA for HIV, hepatitis profile (HBsAg, HBeAg, HBcIgM, HDV) HCV-RIBA, VDRL testing.

Achievements:

Developed and validated in-house diagnostic assays for virus infections.Identified signal transduction pathways with potential implication in virus control.Cloned, expressed and purified several proteins even soluble forms.Prepared monoclonal, polyclonal antibodies and characterized several glycoproteins.Provided insights into transcriptional modulation by using Microarray and data mining.Designed gel-shift and other tools for easy screening of antiviral drug resistance and protein-DNAinteractions.Characterized mechanisms of drug resistance at genome level for herpes viruses.Strong publication record that include manuscripts in peer-reviewed journals.Excellent organizational and teamwork skills, coordinated a group of researchers.Adept in trouble shooting and consistently offered creative solutions for colleagues.

Experience:Current Position:

Managing lab budget, grants, organize and lead projects in a molecular microbiology laboratory. Introduce novel concepts, experiment design, analysis of data, publications, representation at scientific meetings, and meeting overall project time lines. Inaddition, coordinate research activities of other postdoctoral fellows and technicians in the lab.The lab’s focus is microbiology, molecular biology and signal transduction.

Postdoctoral Research in gene regulation:

Postdoctoral fellowship (2002-2003) at Thrombo-Modulin Lab, Blood Research Institute, Milwaukee, Wisconsin, USA. Research activitiesincluded Cloning, expression and characterization of genes involved in Diabetes. Postdoctoralfellowship (2003-2005) at Gastero-Intestinal Research, Massachusetts General Hospital, HarvardMedical School, Boston, MA, USA. Research activities include the studies on mechanism of how fat-soluble (lipophilic) vitamins like Vitamin D and Vitamin A regulate the expression of enterocyte differentiation marker gene, Intestinal alkaline phosphatase (IAP).

PhD Research:

“Studies on protection and immunity induced by

Mycobacterium habana

against

Mycobacterium tuberculosis

in experimental tuberculosis”.

MS Research:

Effect of pyridoxine on the activities of glutamate dehydrogenase, acetylcholinesterase and on neurotransmitters during the growth of

Macrobrachium rosenbergii.

Detailed Chronology:Assistant Professor and Lab Director, Central Research Laboratories, Dept. of Botany &Microbiology, King Saud University, Saudi Arabia. (2005-Present)

Set up the Central Core Lab for proteomics & genomics.

Performed bacterial isolation, identification, antibiotic sensitivity assessmentPerformed ELISA for RSV, EMCV, HCV, HBVManaged personnel, inventory, equipments, clinical meetings, teaching/training graduate andundergraduate studentsTranscriptional profiling of RSV infections using micro array analysis.Developed number of methodologies for anti-viral drug efficacy evaluations.Coordinated a group of 6 researchersIdentification of novel Signal transduction pathway with implication in virus control.Optimized cell-culture for virus isolation, identification, and serotyping, PCR, IFA assays for thelab diagnosis of herpes simplex virus (HSV) infections.Cloned, expressed (GST/His Tag proteins in baculovirus mediated Sf-9/High Five cells), column purified, raised antibodies to characterize glycoproteins gH, gL, gM, gB of RSVDeveloped tear secretory IgA ELISA for the diagnosis of herpes infections.Developed a novel assay for drug resistance of Herpes using PCR-SSCP and gel shift.Developed single tube multiplex PCR for respiratory virusesStudied the role of L-carnitine and Black seed as antiproliferative agent in EMCV infectedWISH cell lines

Post-Doctoral Fellow, Blood Research Institute, Wisconsin (2002-2003) and MassachusettsGeneral Hospital, Harvard Medical School, Boston (2003-2005)

Member, to identify the role of immune-associated nucleotide (IAN) gene in Type I diabetes in ratmodelLead member, identification of human intestinal alkaline phosphatase gene as a gut defense factor Member, created human Intestinal alkaline phosphatase knock-out mouseMember, created human intestinal cells overexpressing Intestinal alkaline phosphatase geneMember, created siRNA to silence human Intestinal alkaline phosphatase geneMember, identified different HDAC inhibitors as potent anti colon cancer agentsPerformed various techniques like: Chromatin Immunoprecipitation Assay, Creating Transgenicmodels, Electrophoretic Mobility Shift Assay, Northwestern blot assay, Multiplex PCR, Real TimePCR, Yeast Two Hybrid Screen Assay, FRET, DNA Footprinting, Microarray, Nuclear Run-on Assay,Fluorescent

In situ

Hybridization, siRNA, Quantum Dot labeling, Transfection,Taught Molecular biology course work for undergraduate students.

PhD Student (CSIR Junior & Senior Research Fellow), Jamia Hamdard University, NewDelhi & at Central Drug Research Institute, Lucknow, India (1996-2002)