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CURRICULUM VITAE
VIKAS KAITHWAS PhD
Molecular Biology and Microbial Physiology Division
CSIR-Institute of Microbial Technology (IMTECH)
Sector 39A, Chandigarh-160036, India
Mobile: + 91-9357393695
Email: [email protected]
PROFESSIONAL and RESEARCH INTERESTS
Challenging position in Life science Industry or Global Intellectual Property Solut ion
Company, Molecular Biology, Cell Biology and Immunology, Genetic engineering, yeast
biology, Protein Science and DNA-Protein Interaction, Signalling, Microbial virulence
studies, Host-Pathogen Interaction
EDUCATION
2009-2014: Doctor of Philosophy (Ph.D.), Biological Sciences, April 2015
Mentor: Dr. Saumya Raychaudhuri,
Senior Scientist, Molecular Biology and Microbial Physiology Division
Institute of Microbial Technology, Chandigarh-160036
Council of Scientific and Industrial Research (CSIR), India
PhD Thesis: “Exploration of Type III Secretion System effectors of
Vibrio parahaemolyticus using Yeast Saccharomyces cerevisiae as a
model system”
2007-2009: Master of Science (M.Sc.), Biotechnology
The Maharaja Sayaji Rao University of Baroda,
Vadodara, India
2 years parallel research project: “Understanding the role of non-
coding RNA in virulence of Pseudomonas aeruginosa”
2004-2007: Bachelor of Science (H), Biotechnology
Bundelkhand University, Jhansi, Uttar Pradesh, India
Training experience during graduation:
“15 days lab training in ICMR-National JALMA Institute for
Leprosy & other Mycobacterial diseases, Agra”
“15 days lab training in Cancer Hospital & Research Institute,
Gwalior”
RESEARCH SKILLS
Immunological and Cell biological techniques:
Western blot analysis for analysing protein expression, stability and signalling
pathways
Yeast gene deletion library screening for the identification of eukaryotic proteins
targeted by bacterial effectors
Microscopy: Morphological imaging through differential interference contrast
(DIC) microscope; Fluorescent microscope and Confocal microscope for
visualizing different fluorescent dyes and fluorescent proteins
FACS for yeast cell cycle analysis
Ultracentrifuges for membrane fractionations
Fluorescent spectrophotometry for actin polymerization assays
Molecular genetics techniques:
Basic cloning techniques (Genomic DNA, RNA and plasmid purification, PCR
amplification, restriction digestion and ligation using variety of enzymes)
Homologous re-combinational cloning
Gene deletion/disruption in bacterial chromosome
Site directed mutagenesis (both conventional (overlapping PCR based) and Kit
based methods): domain deletion and sequential truncation studies, alanine
scanning, protein tagging with different tags such as GFP, mCherry, FLAG to
visualize protein of interest in vivo and in vitro
DNA sequencing and chromatogram analysis
Microbiological techniques:
Microorganism culturing (bacteria, fungus and yeast) and cryopreservation
Cell line handling
Growth Kinetics and growth inhibition assays
Vibrio co-culturing studies with Candida and other fungi
Bacterial culture supernatant assays
Protein biochemistry:
Heterologous protein expression in E. coli
SDS electrophoresis (Coomassie and Silver staining)
Protein purification using affinity chromatography (Ni-NTA for His tagged
proteins)
Protein estimation
Biochemical assays such as actin polymerization assay, protease assay
Basic bioinformatics techniques:
Primer designing
Understanding of bioinformatics tools for day-to-day sequential analysis using
BLAST and ClustalW, etc.
Knowledge of routine softwares, such as Sigmaplot, Micrococal Origin, Adobe
photoshop, End-Note, etc.
Structural analysis of proteins by different protein visualization softwares like
PYMol, Swiss PDB Viewer, etc.
Protein structure prediction, homology modelling, Protein-protein docking (online
servers like tasser, HEX)
Good Laboratory Practices:
Independent planning and execution of experiments
Daily documentation of experimental plans, protocols and data
Analysis and interpretation of experimental data; scientific discussion in lab
meetings, journal club presentations
Writing manuscripts and other technical reports independently
AWARDS, FELLOWSHIPS AND ACADEMIC ACHIEVEMENTS
2011 to 2014: Senior Research Fellowship
Awarded by Council of Scientific and Industrial Research,
Govt. of India
2009 to 2011: Junior Research Fellowship
Awarded by Council of Scientific and Industrial Research,
Govt. of India
Dec. 2008 and June 2009: Qualified National Eligibility Test (CSIR-NET)
Conducted by Council of Scientific and Industrial Research,
(CSIR), Govt. of India.
2007-2009: Awarded scholarship by Department of Biotechnology,
New Delhi, Govt. of India
(During M.Sc. Biotechnology at Maharaja Sayaji Rao
University of Baroda, Vadodara, India)
2007: Qualified Combined entrance examination (CEEB) conducted
by Jawaharlal Nehru University (JNU) for admission to
M.Sc Biotechnology programme academic session
CONFERENCES/ WORKSHOPS ATTENDED
2013: Attended the “8th International Conference on Yeast Biology”
Organized by CSIR Institute of Microbial Technology (IMTECH),
Chandigarh, India.
2011: Attended the “7th International Conference on Yeast Biology”
Organized by Indian Institute of Technology, Mumbai, India.
2010: Attended an international conference on “Understanding and Managing
Pathogenic Microbes (UMPM-2010)”
Organized by CSIR Institute of Microbial Technology (IMTECH),
Chandigarh, India.
2006: Attended National Conference on “Immunology in Health and Diseases”
Organised by Chhatrapati Shahu Ji Maharaj University, Kanpur.
2006: Attended National Symposium on “Recent Trends in Molecular Biology”
Organised by Cancer Hospital and Research Institute, Gwalior.
ORGANIZATIONAL SKILLS
Worked as class representative in graduation and post graduation responsible for managing
lecture and lab experiment schedules, seminars and most importantly working as a
communication bridge between faculty and students
PUBLICATIONS
Ranjana Tripathi1, Vikas Kaithwas
1, Chetna Dureja
1, Saumya Raychaudhuri
* 2013.
Alanine-scanning mutagenesis of WH2 domains of VopF reveals residues important
for conferring lethality in a Saccharomyces cerevisiae model, Gene 525: 116–123
(1 = equal contributors)
Vikas Kaithwas, Prasant K Dhaure, Leela Krishna Bangkapalli and Saumya
Raychaudhuri*. Functional characterization of VopR by exploiting budding yeast as a
model system (Manuscript under preparation)
PERSONAL DETAILS
Date of Birth: 26th September, 1986
Place of birth: Jhansi, Uttar Pradesh, India
Nationality: Indian
Gender: Male
Marital Status: Un-married
BRIEF SYNOPSIS OF RESEARCH (2009-2014)
Vibrio parahaemolyticus is a gram negative, halophilic bacterium ubiquitously found in
marine and estuarine environments. The bacterium is often associated with food-borne
gastroenteritis and traveler’s diarrhea, thereby becoming an increasing health concern across
the globe. Genome sequencing of a virulent strain of a KP-positive V. parahaemolyticus
strain RIMD 2210633 reveals the armamentarium of pathogenic determinants containing
thermostable direct hemolysins (TDH) and two type-III secretion systems (T3SS). It has been
demonstrated that T3SS1 is involved in cytotoxicity whereas T3SS2 is responsible for
enterotoxicity and cytotoxicity of V. parahaemolyticus (Ono et al., 2009, Caburlotto et al.,
2010, Kodama et al., 2007). By employing tissue culture and animal model system, a myriad
of T3SS effector proteins have been identified from V. parahaemolyticus. These effectors are
not only unique in terms of their structural and functional aspects but also target and
modulate disparate/distinct cellular processes to evade host surveillance system (Broberg et
al., 2011). In recent time, Saccharomyces cerevisiae, the budding yeast has gained much
attention as a non-mammalian model system to identify and evaluate functionality of diverse
arrays of virulence factors not only for its easy cultivation but also presence of cellular
pathways which are well conserved in mammalian systems (Valdivia, 2004; Curak et al.,
2009).
Until now, functional aspect of VopR remains elusive or are poorly explored (Broberg et al.,
2011). In order to gain insight, we have chosen to examine its functionality in budding yeast
model system. Herein, our results clearly evidenced that ectopic expression of VopR confers
discernible lethality in S. cerevisiae. Sequential protein truncation analysis identifies critical
regions in the molecule contributing to its toxicity in yeast model system and membrane
localization. We are conducting functional screening of yeast deletion library against VopR
to identify intracellular targets. Furthermore, VopR homologue of V. alginolyticus
homologue has also been identified and examined in yeast model.
VopF, the type III effector molecule, has been implicated in the pathogenesis of non-O1, non-
O139 strains of V. cholerae. It is a protein of 530 amino acids, comprises of one formin
homology 1-like (FH1-like) domain and three WASP homology 2 (WH2) domains. Previous
works have demonstrated that WH2 domains are crucial for VopF function as a modulator of
cellular actin homeostasis. Furthermore, domain deletion analysis also suggests that VopF
variant constituted with only WH2 domain 3 is more efficient in restricting the growth of
budding yeast than its congeners containing either only domain 1 or domain 2. Interestingly,
a good degree of sequence diversity is present within each WH2 domain of VopF. In order to
ascertain the importance of different amino acids in each WH2 domain, a systemic alanine
scanning mutagenesis was employed. Using a yeast model system, the alanine derivatives of
each amino acid of WH2 domain 1 and 3 of VopF were examined for growth restricting
activity. Taken together, our mutagenesis results reveal the identification of critical residues
of WH2 domain 1 and 3 of VopF.
Additionally, I have got an opportunity to investigate the interaction of Vibrio spp. with
Candida albicans and other fungi. In parallel, I have also gained knowledge regarding some
other projects going on in our lab including functional role of quorum sensing master
regulatory proteins, in the context of DNA- Protein interaction (gene regulation).
REFERENCES
Saumya Raychaudhuri, Ph.D.
Ph.D. Supervisor (Scientist E)
Molecular biology Divison
IMTECH, Chandigarh-160036, India
E-mail: [email protected]
[email protected] Phone: +91-172-6665256,
Mobile: +91-9417727654
Dr. Ashish Senior Scientist,
Protein Science and Engineering Division
CSIR-Institute of Microbial Technology
Sec. 39A, Chandigarh-160 036, India
Phone: +91172666472 (Lab)
E-mail: [email protected]
Alok Kumar Mondal, Ph.D.
Professor
School of Life Sciences
Jawaharlal Nehru University
New Mehrauli Road, New Delhi-110067
Email: [email protected] Phone: +91-11-26742676
K. Ganesan, Ph.D.
Scientist F
Institute of Microbial Technology
(Council of Scientific and Industrial Research)
Sector 39A, Chandigarh-160036, India
Email: [email protected] Phone: +91-172-6665305,
Mobile: +91-9464395061
