Ahmad Aktsar Roskiana, Kaewpungsup Pornjira, Khorattanakulchai Narach, Rattanapisit Kaewta, Pavasant Prasit, Phoolcharoen Waranyoo
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Laboratory of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Universitas Muslim Indonesia, Makassar, Indonesia.
Biotechnol Rep (Amst). 2019 May 23;23:e00348. doi: 10.1016/j.btre.2019.e00348. eCollection 2019 Sep.
The study aimed to produce recombinant human dentin matrix protein 1 (DMP1) and to test, whether the recombinant DMP1 produced in 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 . 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 can induce the osteogenic differentiation of human PDL cells, leading to improved tooth repair and regeneration.
该研究旨在生产重组人牙本质基质蛋白1(DMP1),并测试在[具体宿主]中产生的重组DMP1是否具有功能活性。使用pET22b质粒构建了一个在其C末端具有多组氨酸序列的编码DMP1蛋白的基因结构,并在[具体宿主]中表达。培养条件的优化使得能够用0.5 mM异丙基β-D-1-硫代半乳糖苷(IPTG)诱导基因表达,并在37°C下培养6小时以生产DMP1重组蛋白。使用镍亲和层析法纯化重组蛋白。检测了DMP1对人牙周膜(PDL)细胞活力、成骨分化和钙化的影响。纯化的DMP1可诱导PDL细胞中与成骨相关基因的表达和钙沉积。这些发现表明,在[具体宿主]中产生的DMP1可诱导人PDL细胞的成骨分化,从而改善牙齿修复和再生。