Improving the Recombinant Protein Expression of Human Galectin-3 in BL21 Bacterial Host System
Praveen Kumar Vemuri *
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Nikhil Reddy Varakala
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Divyanshu Dhakate
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Tripura Ravavarapu
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Faina Philberta Dumpala
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Susmita Sri Muddana
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Haritha Bommepalli
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
Spurthi Modiboyana
Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Andhra Pradesh, India.
*Author to whom correspondence should be addressed.
Abstract
Background: Regardless of the broad explore in the territory of glycobiology concerning structure and capacity of glycans, lectins and glycosylation forms, numerous viewpoints are still left unexplored.
Aim: In this study, we analyzed the effect of shuttle vector on the secretion of human galectin recombinant protein.
Methods: The galectin was expressed in E. coli BL21 by growing the bacterial culture in SOC medium and purified by nickel-based affinity chromatography due to its His-tag.
Results: After cell lysis the protein was identified as a single 29 KDa band by 12% SDS-PAGE. Conclusion: Characterization studies clearly revealed that the purified protein was indeed galectin 3.
Keywords: Galectin-3, glycobiology, BL21, glycan, cloning, expression.