Volume 15, Issue 35, January - June, 2021

Isolation, cloning and transformation of glucan binding protein (GBP) gene

Ravikumar Patil HS^1♦, Vasantha Naik T², Vijay Avin BR³, Sayeswara HA⁴

¹Department of Biotechnology, G.M. Institute of Technology, Davangere-577066, Karnataka, India
²Department of Botany, D.R.M. Science College, Davangere University, Davangere-577066, Karnataka, India
³Department of P.G. Studies and Research in Biotechnology, Sahyadri Science College (Autonomous), Kuvempu University, Shivamogga-577203, Karnataka, India
⁴Department of Zoology, Sahyadri Science College (Autonomous), Kuvempu University, Shivamogga-577203, Karnataka, India

^♦Corresponding author
Department of Biotechnology, G.M. Institute of Technology, Davangere-577066, Karnataka, India, E-mail: patil_varuni@reddifmail.com

ABSTRACT

This work was to isolate the glucan binding protein (gbp) gene from Streptococcus mutans, it possesses at least four glucan binding proteins GbpA, GbpB, GbpC, and GbpD, each of which appears to be immunologically and biochemically distinct using various processes. Firstly, the organism was cultured using the brain heart infusion broth media. Later the isolation of the gene was carried out using several processes such as the ones beginning from electrophoresis, isolation of the total protein and genomic DNA etc. Then, cloning was carried out using Escherichia coli as the host and the competent cells were prepared. In the second part, the competent cells were cultured in a media containing IPTG, X-gal, ampicillin and the transformed cells were differentiated from the recombinant cells by the color developed by the colonies. In later stages, the gbp gene that was isolated was sequenced and the query sequence that was obtained was analyzed using bioinformatics tools like BLAST and the similarity search was carried out in order to find and derive its various applications

Keywords: Streptococcus mutans, glucan binding protein (gbp), PCR, competent cells

Drug Discovery, 2021, 15(35), 16-22

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