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Cloning, Expression and Purification of Recombinant Human Galectin-3 by Using pPUZZLE Shuttle Vector in Bacterial Strain (DE3) Host System

Author(s): Vemuri Praveen Kumar | Veeravalli Suryanarayana | Poda Sudhakar | Krothapalli Raja Surya Sambasiva Rao

Journal: Advanced Biotech
ISSN 0973-0109

Volume: 13;
Issue: 01;
Start page: 09;
Date: 2013;
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Keywords: Galectin-3 | pPUZZLE vector | Epx1 | MFα1pp | DE3 bacterial strain

Galectin-3, an 29KDa protein is a unique member of the growing family of β-galactoside binding lectins, contains a single carbohydrate recognition domain and a glycine rich N-terminal domain, through which it can form oligomers and functions to cross-link both carbohydrate and non-carbohydrate ligands. Galectin-3 is widely expressed in adult tissues particularly on and secreted by activated macrophages, onocytes and adipocytes. Galectin-3 has been implicated in many facets of the inflammatory response including neutrophil and macrophage activation and function. In this work, the novel pPUZZLE vector is successfully used for recombinant protein production in BL21 (DE3) strain. The application of a strong promoter (e.g. extracellular protein X; Epx1) and efficient secretion signals (alpha mating factor secretion leader; MFα1pp) as well as proper folding and post- translational modifications are beneficial to produce human galectin-3.

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