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Cloning and production of the monomer form of recombinant streptavidin and appraisal of it`s binding affinity to biotin
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Elham Didevara   , Abdo alrahim Sadeghi , Hamid Abtahi |
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Abstract: (4760 Views) |
| Introduction: Streptovidine is a homo troterameric protein that has a high biotin tendency to use in biotechnology and cellular studies. But its tetramer form interferes in some Measurements. Therefore, in this study, with a deletion mutation, a form of Streptovidine monomer was produced.
Materials and Methods: Firstly, the streptavidin nucleotide sequence was taken from NCBI site and after mutation in the sequence of the gene was optimized for expression in E.coli bacteria. Correspondingly, in order to clone the gene, E. coli DH5α strain and to express the streptavidin gene E. coli BL21 (DE3) pLysS were used. Purification of protein was performed with Ni-NTA kit and by HABA dye, the tendency of binding of streptavidin to biotin was measured.
Results: Streptovidine monomer form was produced and purified in E.coli BL21 (DE3) pLysS expression host. The recombinant Streptovidine monomer has a tendency for binding to biotin.
Conclusion: By creating a mutation in the streptavidin peptide sequence, it can be obtained in the form of a monomer that has a tendency for binding to the biotin |
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| Keywords: Streptavidin, Cloning, Molecular, Escherichia coli |
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Full-Text [PDF 746 kb]
(1806 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2017/01/16 | Accepted: 2017/11/15 | Published: 2018/01/1
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