Expression and Purification of Recombinant Human Amelogenin Using the Prokaryotic Expression System
- 10.2991/amsee-16.2016.15How to use a DOI?
- recombinant huamn amelogenin; E. coli BL21 cells; purification; SDS-PAGE
The amelogenins (Am) constitute about 90% of the enamel matrix proteins and play a critical role in enamel biomineralization. This study aimed to express and characterize recombinant human amelogenin (rAm) protein in the prokaryotic expression system in quantities sufficient for structural and functional research. Human cDNA coding for a 175 amino acid amelogenin protein was subcloned into the pET-28a(+) vector, this system adds a hexa-histidine tag to N-terminal amino acid of the expressed protein, enabling effective three steps purification by Ni2+-NTA affinity chromatography, an anion-exchange resource Q packed column , and Superdex 200 10/300 GL gel filtration resin. The recombinant protein was expressed in E. coli BL21 cells and the yield of purification his-tagged human amelogenin was up to 19mg/L culture. Recombinant human amelogenin (rAm) was characterized by SDS-PAGE and Coomassie brilliant blue R-250. Production of significant amounts of pure, full-length amelogenin opened up the possibility to investigate novel functions of amelogenin. Their chemical structure, self- assembly and amelogenin-mineral interactions play a key role in the regulation of enamel mineralization process.
- © 2016, the Authors. Published by Atlantis Press.
- Open Access
- This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).
Cite this article
TY - CONF AU - Xiaoyun Feng AU - Jing Yao AU - Qin Du AU - Xiaohua Ren AU - Kun Tian PY - 2016/04 DA - 2016/04 TI - Expression and Purification of Recombinant Human Amelogenin Using the Prokaryotic Expression System BT - Proceedings of the 2016 International Conference on Advanced Materials Science and Environmental Engineering PB - Atlantis Press SP - 51 EP - 54 SN - 2352-5401 UR - https://doi.org/10.2991/amsee-16.2016.15 DO - 10.2991/amsee-16.2016.15 ID - Feng2016/04 ER -