摘要
目的 运用基因重组的方法 ,对人TGF β1的进行基因克隆 ,在大肠杆菌中表达 ,并进行重组TGF β1修复骨缺损的实验研究。方法 以pBV2 2 0 作为原核细胞表达载体 ,将TGF β1cDNA片段定向重组到pBV2 2 0 的多克隆位点上 ,构建原核细胞表达质粒pTGF β1并转化至大肠杆菌中进行温度诱导表达。在兔顶骨两侧各制造直径为 10mm的骨缺损 ,左侧缺损植入纤维蛋白加重组TGF β110 0ug作为实验组 ,右侧缺损单纯植入纤维蛋白作为对照组 ,术后进行组织学和平均灰重测定。结果 表达产物经SDS PAGE分析 ,该表达体系可高效表达TGF β1,其表达产物占菌体可溶性蛋白的 2 3% ,用NRK细胞检测本研究生产的TGF β1具有该蛋白的野生活性。实验组缺损区组织学及平均灰重明显优于对照组 (P <0 0 5 )。结论 TGF β1原核表达体系的建立为TGF β1的生产提供了高效的表达工程菌 ,重组TGF
Objective [HT5”SS]Using the method of gene recombination to express TGF β1 in Escherichia Coli and to determine its ability of repairing bone defects.[HT5”H]Methods [HT5”SS]TGF β 1 cDNA fragment was inserted into pBV 220 , a prokaryotic expression vector,the recombinant plasmid was transformed into E.coli to establish the prokaryotic expression system and induced to express encoded protein.The experimental models were made by drilling lcm circular defects in bilateral parietal bones of the rabbit.The left defect was used as a carrier of filled with TGF β 1 composite and the right side was filled with fibrin.Four weeks after operation,histological changes were observed and ash weight measured.[HT5”H]Results [HT5”SS]The system can express efficiently TGF β 1,which was 23 percent of the total bacterial soluble protein.The osteogenesis of TGF β 1 and fibrin group surpassed obviously that of fibrin group.[HT5”H]Conclusion [HT5”SS]The system is able to stimulate obviously the repair of bone defects.
出处
《中国骨伤》
CAS
2000年第12期715-717,共3页
China Journal of Orthopaedics and Traumatology
