Recombinant human dentin matrix protein 1 (DMP1) induces the osteogenic differentiation of human periodontal ligament cells

Autor: Aktsar Roskiana Ahmad, Pornjira Kaewpungsup, Narach Khorattanakulchai, Kaewta Rattanapisit, Prasit Pavasant, Waranyoo Phoolcharoen
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Biotechnology Reports, Vol 23, Iss , Pp - (2019)
Druh dokumentu: article
ISSN: 2215-017X
DOI: 10.1016/j.btre.2019.e00348
Popis: The study aimed to produce recombinant human dentin matrix protein 1 (DMP1) and to test, whether the recombinant DMP1 produced in Escherichia coli possesses functional activity. A gene construction comprising a gene encoding for DMP1 protein with polyhistidine sequence at its C-terminus was created using the pET22b plasmid and expressed in E. coli. The optimization of cultivation conditions has enabled the induction of the gene expression with 0.5 mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and DMP1 recombinant protein production at 37 °C for 6 h. The recombinant protein was purified using Ni affinity chromatography. DMP1 influence on the viability, osteogenic differentiation and calcification of human periodontal ligament (PDL) cells was examined. The purified DMP1 could induce the expression of osteogenesis related genes and calcium deposition in PDL cells. These findings indicate that DMP1 produced in E. coli can induce the osteogenic differentiation of human PDL cells, leading to improved tooth repair and regeneration. Keywords: Dentin matrix protein 1 (DMP1), Prokaryotic expression, E. coli, Purification
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