Master of Biotechnology

SUMMARY:

Recently graduated from Texas A&M University with Professional Science Masters in Biotechnology. At present, actively seeking full time job opportunities to gain industrial exposure and utilize my skills. Two year research experience in molecular biology, cell culture and downstream processing. Quick learner with good communication and management skills. With a strong sense of team spirit, I look forward to be a valuable addition to your organization.

TECHNIQUES:

SDS PAGE

• Ran gels for qualitative analysis of recombinant proinsulin and mature insulin expression

• To analyze and compare the size of mammalian and avian immunoglobulin

ELISA (Immunoassay)

• Sandwich ELISA for goat-antimouse-IgG using the peroxidase-hydrogen peroxide detection system to assay the concentration of mouse IgG in the sample

• Competitive ELISA to detect the presence of anti-thyroglobulin antibodies

Protein Assays

• BCA kit to quantify the expressed crude proinsulin and to compare the protein expression after refolding the disulfide bonds in proinsulin

• Malachite Green activity assay to determine the concentration of active MurA protein based on the release of inorganic phosphate

FPLC

• To purify the recombinant mature insulin using AKTA-FPLC system in sodium citrate buffers from a concentration of 0M to 0.1M

Affinity Chromatography

• Purification of sfGFP linked to unnatural amino acids using nickel and cobalt affinity columns

• Purification of rabbit IgG using Protein A affinity chromatography with a CNBr activated sepharose matrix immobilized with protein A

Western Blot

• Carried out SDS PAGE followed by western blot to detect the chicken growth hormone in the pituitary extract

• Used the blot for the detection of MurA protein

PCR

• Performed big dye sequencing PCR to check for site specific mutations

• Carried out diagnostic PCR for site directed mutagenesis of proinsulin gene and to amplify cloning vector pBS containing the MurA gene

Bacterial Gene Cloning

• Cloned the MurA gene into pBS vector using restriction digestion, ligation, transformed it into XL-1 blue cells, screened transformants using blue-white screening, DNA extraction using Qiagen kit followed by plasmid DNA analysis using NanoDrop spectrophotometer

• Molecular cloning of proinsulin gene into pEVOL vector and introducing single amino acid substitutions within the proinsulin gene

Vector NTI

• Designed forward and reverse primers for site directed mutagenesis to create insulin analogs such as lispro, aspart, glulisine and glargine

• Sequence analysis of the proinsulin to confirm the presence of desired mutation using Vector NTI and Contig Express

Cell Culture

• Aseptic techniques, maintaining, passaging and cryopreservation of cell lines

Computer Skills

• Microsoft Office Suite (Word, Excel, PowerPoint), EndNote for managing references

WORK EXPERIENCE:

Department of Chemistry, Texas A&M University, College Station, Texas

Graduate Research Assistant | (August 2011- May 2012)

Responsible for carrying out independent research in assigned projects

Worked towards improving insulin yield in bacteria and successfully cloned insulin analogs

Prepared and autoclaved nutrient media and buffers for the lab

Maintained cell lines for research group members and organized lab inventory

Supervised and trained undergraduate students in molecular biology techniques

Interaction with research advisor on a daily basis to set targets and work towards achieving them

Weekly research presentations to the group and positive contributions to the discussions

Research Intern | (May 2011-July 2011)

Carried out research under supervision of research group member

Project involved comparing expression levels of four non canonical amino acids by protein expression, isolation, purification and quantification

SDS PAGE gels were run for a qualitative analysis

Responsible for autoclaving and discarding bio hazardous waste

Prepared media, buffers and autoclaved glassware and pipet tips

Molecular and Microbial Pathogenesis Lab, College of Medicine, Texas A&M University

Lab Research Assistant | (Sept 2010-April 2011)

Assisted in research to study the molecular mechanisms employed by C. burnetti for evading host defenses

In-charge of using recombinant DNA techniques, protein expression and analysis methods

Responsible for carrying out a project to create gene mutation libraries

Advanced Diploma in Genetic Engineering from JIVAS (Jain Institute of Vocational and Advanced Studies)

Trainee | (Jan 2009-Feb 2009)

Molecular Biology: DNA Fingerprinting (RAPD, RFLP), hybridization Techniques (Northern, Southern and Western Blotting), ELISA, electrophoresis (agarose and SDS-PAGE), DNA isolation (Genomic and Plasmid), conjugation, transformation and transduction

Downstream: Chromatographic Techniques (Affinity, Gel Filtration and Ion Exchange Chromatography)

EDUCATION:

Professional Science Masters (PSM) | Biotechnology with Minor in Business| May 2012

Texas A&M University, College Station

Relevant Course work

Molecular Genetics Lab | Jan 2011-May 2011

Start to finish project involving molecular cloning and protein expression using restriction-digestion, isolation and analysis of nucleic acids, ligation, transformation, DNA sequencing, protein expression and purification

Immunology Lab | Aug 2011-Dec 2011

Sandwich ELISA, Hemagglutination, Immunochemical and Immunofluorescent techniques, Ouchterlony double diffusion

B.E | Biotechnology | May 2010

R. V. College of Engineering, Bangalore, India

Relevant course work

Genetic engineering, Enzyme technology, Molecular biology, Animal biotechnology, Immunology, Upstream and Downstream processing lab, Food biotechnology and Project management

ACADEMIC PROJECTS:

Cloning of insulin analogs: Insulin lispro, aspart, glargine and glulisine | (Jan 2012-May 2012)

Primers were designed using Vector NTI

Carried out site directed mutagenesis using “Quik Change” protocol to incorporate desired mutations in the A and B chains of insulin

Carried out restriction digestion, ligation, transformation, colony PCR and plasmid minipreps

Performed sequence PCR and confirmed incorporation of mutations in all sequences

Successfully produced insulin analogs, with each analog differing in properties such as rate of absorption, peak times and retention time in blood

Genetic incorporation of non-canonical amino acids in insulin to expand the genetic code | (Aug 2011 – Dec 2012)

E. coli cells were co-transformed with two plasmids. Proinsulin gene mutated to contain a stop codon in place of lysine at 29th position of B chain and linked to sfGFP

Expression of proinsulin gene in 200 ml LB media, induced with arabinose, IPTG and N-Boc-L-lysine

N-Boc-L-lysine incorporated in place of the stop codon. This prevents lysine recognition by trypsin and subsequent cleavage

Isolation of insulin by cell lysis using sonicator

Qualitative analysis of protein by running SDS PAGE gel

Refolding of insulin by to aid in formation of disulfide bonds

Carried out quantitative analysis of protein using the BCA assay to determine protein concentration

Digestion of proinsulin using trypsin to form mature insulin

The formation of mature insulin was confirmed using SDS PAGE

Genetic Incorporation of non-canonical amino acids in proteins to expand the genetic code | (May 2011-July 2011).

Creating gene mutation libraries using Himar1 transposon | (Jan 2011-April 2011)

Production of Biodiesel from Algae | (Jan 2010-May 2010)

Production of Bioethanol from lignocellulosic materials using Phanerochaete chrysosporium | (Fall 2008)

SEMINAR PRESENTATIONS:

Production of Biodiesel from Microalgae | May 2010

Phyto-Pharmaceuticals and its role in topical pain relief | April 2010

EXTRA-CURRICULAR ACTIVITIES:

Team Leader for College based Social Club, RAAG-The Youth Club of RVCE | Organized visit to orphanages

Core committee member for Biotechnology Department Fest, HELIX | Organizing and managing department fests

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