Lokesh Bhushan Scientist- Drug Discovery
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I did a Ph.D. in Biochemistry from the Indian Institute of Science, Bangalore, India. I have almost seven years of experience in academic research and two years of experience in the industry. I am currently working as a scientist at InveniAI (a subsidiary of BioXcel Corporation), Bangalore, India. Teaching and mentoring to the students is my hobby. I have helped many students and colleagues to understand the basic concept of biology. My asset is to explain complex biological concepts in simple language using an appropriate analogy.

Question
Let's assume you received two DNA samples: DNA A and DNA B. One of DNA contains promoter sequences. Design an experiment that allows you to find out which DNA samples contain the promoter sequences. Assume you have access to all the required reagents (like primers, polymerase enzyme, dNTP, plasmids, etc.) and resources to execute the experiments.

Answer:

Luciferase reporter assay is the most accepted tool to identify and characterized the promoter sequence.
Both DNA samples will be clone in pGL3 Luciferase reporter vectors, individually.
A reporter constructs containing DNA A named as pGL3-A whereas, reporter construct containing DNA B named as pGL3-B.

Mammalian cells will be culture in 3 wells of six-well plate up to 70%–80% confluence. One well will be transfected with plasmid pGL3-A and
the second well will be transfected with plasmid pGL3-B and third well will be transfected with vector alone pGL3 (served as a negative control).
After incubation for a desired time, luciferase activity will be measured in each transfected sample.
A DNA sample that contains the promoter sequence would show luciferase activity, and a DNA sequence that does not contain promoter sequence would not show luciferase activity.

For example, if we detect luciferase activity in a sample transfected with pGL3-A plasmid. It implies that DNA A contains a promoter sequence.

Subjects

  • Biological Science Grade 9-Masters/Postgraduate

  • Biochemistry Grade 11-Masters/Postgraduate

  • Microbiology Grade 12-Masters/Postgraduate

  • Cell Biology Grade 11-Masters/Postgraduate

  • Cell and Molecular Biology Grade 12-Masters/Postgraduate

Experience

  • Manager Scientific Analytics (Dec, 2019–Present) at InveniAI (Subsidiary of BioXcel Corporation)
    Drug-Repurpsing
  • Senior Research Investigator (Jul, 2018–Dec, 2019) at Syngene Int. Ltd. Bangalore India
  • Scientist (Jul, 2017–Jun, 2018) at Shilpa Medicare Ltd. (Formerly Navya Biological Pvt. Ltd.)
  • Postdoctoral Fellow (Jan, 2017–Jul, 2017) at National center for Biological science India
  • Research Associate (Jun, 2015–Jun, 2016) at Midwestern University USA
  • Research Fellow (Jan, 2014–May, 2015) at Cincinnati Children’s Hospital Medical Center USA
  • Postdoctoral Fellow (Jan, 2011–Jan, 2014) at Western University of Health Sciences USA

Education

  • PhD (Jul, 2005–Dec, 2010) from Indian institute of science, Bengaluru
  • MS (Jul, 2002–Jun, 2005) from Indian institute of science, Bengaluru

Fee details

    ₹600–700/hour (US$7.19–8.38/hour)

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