Construction of A Protein Purification System Mediated by Split Intein

doi:10.3969/j.issn.1673-1689.2019.10.019

Home > Archive>Volume 38, Issue 10, 2019 >135-143. DOI:10.3969/j.issn.1673-1689.2019.10.019

Construction of A Protein Purification System Mediated by Split Intein
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                        10.3969/j.issn.1673-1689.2019.10.019
                    
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Molecular biology techniques are used to modify the N（I_N） and C（IC） fragment of Npu DnaE（Nostoc punctiforme） intein. An affinity chromatography medium with I_N as the affinity ligand and a fusion protein expression system with IC as self-cleavage affinity tag were constructed. Finally，a protein purification system mediated by split intein was developed. The effect of steric hindrance on the C-terminal cleavage rate of fusion protein was studied by constructing 3 kinds of IN affinity ligands and 2 kinds of IC-GFP fusion proteins. Through the combination of in vitro cleavage and Zn²⁺ inhibition experiment，we found the best combination of F and the new Zn binding site. The I_N affinity chromatography media were successfully prepared by using cysteine at the C-terminal of N3 affinity ligands. The purification effect of it was studied，and the high purity GFP protein was obtained successfully.

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WANG Yujun, WANG Shujing, DU Yexing, FENG Lili, XIA Haifeng. Construction of A Protein Purification System Mediated by Split Intein[J]. Journal of Food Science and Biotechnology,2019,38(10):135-143.

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