摘要
目的:利用基因工程技术生产人骨形成蛋白(hBMP),为其诱骨机理及临床应用研究提供前提条件。方法:作者将hBMP2cDNA3’端732bp片段,克隆入大肠杆菌表达载体pGEX-2T,经IPTG诱导后,表达产物存在于包涵体中。将其纯化复性后,进行SDS-PAGE和FPLC分析。并作小鼠体内异位诱骨活性检测。结果:表达的hBMP2与GST的融合蛋白Mr=51ku,占菌体总蛋白的30%。纯化、复性后,FPLC分析示尖而窄的洗脱峰。SDS-PAGE呈一条带。在小鼠肌肉中可诱导软骨及骨组织生成。结论:hBMP2在大肠杆菌中的表达产物复性后纯度较高,且具有诱骨活性。
Objective:By way of genetic engineering production of human bone
morphogenetic protein(hBMP2),it will offer the basis for the clarification of its bone inductive
mechanism and clinical application. Methods:3’terminal fragment of hBMP2 cDNA with size of
732 bp was cloned into pGEX+2T expressionvector and induced by IPTG,the expressed product
existed in a form of inclusion body.After purification andrenaturation of rhBMP2,its bone
inductive activity was assayed in mouse heterotopically in vivo. Results:The fusion protein of
GST with hBMP2 is 51 ku in size. After purification and renaturation,FPLC analysisshowed a
sharp and narrow elution peak.The result on SDS-PAGE appeared in vivo one band.The
histolog-ic assay displayed that rhBMP2 can induce the formation of cartilage and bone tissue.
Conclusion:TherhBMP2 expressed in E,coli was able to induce bone formation after
renaturation of inclusion body. The re-natured protein had a relatively high purity.
出处
《第四军医大学学报》
1995年第6期429-431,共3页
Journal of the Fourth Military Medical University
关键词
人骨形成蛋白
基因表达
大肠杆菌
包涵体
human bone morphogenetic protein
gene expression
E. coli
inclusion body