Summer Research Fellowship Programme of India's Science Academies

Subcloning of IQGAP2 cds in pcDNA3.1 Vector

Charu Jain

University of Hyderabad 

Dr. Manjusha Dixit

Reader F, School of Biological Sciences, National Institute of Science Education and Research, Jatni, District, Khurda, Odisha


Gene cloning is a very convenient molecular biology technique to amplify a fragment of DNA and its expression in a suitable host. Here, we tried to subclone IQGAP2 gene from a cloning vector pCMV6 (4.9kb) to a mammalian expression vector pcDNA3.1. IQGAP2 gene product is known to interact with components of the cytoskeleton and to act as a tumor suppressor and also has been found to play a role in regulating innate antiviral response. We followed PCR based method of subcloning and the cloning was confirmed by colony PCR. However, it needs further validation by sequencing and then the construct can be transfected into a mammalian cell line to study its function.

Keywords: IQGAP2, Subcloning, PCR, Restriction digestion, Transformation, Selection, Sequencing.

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