Expression, Purification of Recombinant Human Thymosin β16 and Applications of Thymosin β16
|Keywords||Thymosin β16 Wound healing Ulp1 SUMO|
The Tβ16 as one of the thymosin beta family members , by anti-inflammatory and immunomodulatory effects , stimulating the proliferation of fibroblasts , promoting keratinized epithelial cells , stimulate endothelial cell migration and promote angiogenesis to promote wound repair . Filter by cloning the subject application of recombinant technology, design to build containing SUMO - Tβ16 fusion of recombinant plasmid SUMO-Tβ16/pET3c of the plasmid was transformed into E. coli BL21 (DE3) , two entirely correct recombinant DNA sequencing engineered bacteria SUMO-Tβ16/pET3c/BL21 . The engineered bacteria continuous passage 100 generations , the indexes were good genetic stability . The restructuring SUMO - Tβ16 Fusion Engineering bacteria induced by 0.8 mmol / L IPTG , 30 ° C culture 6h after SDS - PAGE electrophoresis for soluble expression . By DEAE anion exchange chromatography → hydrophobic chromatography Cut → → SUMO proteases ( Ulp1 ) gel filtration chromatography → purity of greater than 95 % and a specific activity of 5.3 × 105IU/mg Tβ16 . In vitro and in vivo biological activity experiments show : Tβ16 promote fibroblast proliferation ; promote the proliferation of rabbit corneal cells ; promote the chick chorioallantoic membrane vascular proliferation ; Tβ16 gel can promote the healing of rabbit skin burns and alkali burn . In summary, the present research results show that Tβ16 by their anti- infection , to promote the proliferation of fibroblasts and promoting the regeneration of blood vessels and other effects on wound repair , initially to determine the Tβ16 effective doses for 5 ~ 10μg / g , Tβ16 drug development . experimental basis .