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Iranian Journal of Biotechnology، جلد ۱۲، شماره ۲، صفحات ۳-۸
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عنوان انگلیسی Expression, Purification and Characterization of Human Recombinant Galectin 3 in Pichia pastoris
چکیده انگلیسی مقاله Background: Over the past century, the areas of genomics, proteomics and lipids have captured the attention of investigators worldwide. Carbohydrates, have recently received increased attention through the expanding field of glycobiology; probably because they are very complex and not encoded in the genome. Objectives: The purpose of this study was to express and purify recombinant human galectin 3via the Pichiapastoris expression system. Materials and Methods:cDNA of human galectin 3 gene was amplified with specific primers and cloned into a pcDNA3.1 vector with His-tag for easier purification using Ni2andchromatography. Furthermore, galectin 3was purified to homogeneity and confirmed using SDS-PAGE and western blotting. Results:The protein band corresponding to 29 kDa was excised from the gels, digested with trypsin and processed for mass spectrometric analysis by Matrix Assisted Laser Desorption/Ionization- Time of Flight Mass Spectroscopy (MALDI-TOF MS), using a Reflex III instrument. Conclusions:Tryptic digest analysis clearly revealed that the purified protein was indeed galectin 3. Similarly, the biological activity of recombinant galectin 3 was confirmed using the hemagglutination inhibition assay.
کلیدواژه‌های انگلیسی مقاله Pichiapastoris,Galectin 3,Lgals3,Glycomics,Hemagglutination
نویسندگان مقاله praveen کومار vemuri | kumar vemuri
department of biotechnology, k l university, guntur district, andhra pradesh, india

suryanarayana veeravalli |
department of applied biology, jigjiga univeristy, ethiopia

نشانی اینترنتی http://www.ijbiotech.com/article_7257_9cfff19709b6aa689d9bd3d146f35284.pdf
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