Dissertation > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)

Expression, Purification of Recombinant Human Thymosin β16 and Applications of Thymosin β16

Author FengXiuPing
Tutor ZhuXun
School Jilin University
Course Immunology
Keywords Thymosin β16 Wound healing Ulp1 SUMO
Type PhD thesis
Year 2009
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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 .

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