摘要
目的用基因工程技术在大肠杆菌中表达人骨形态发生蛋白２（ｈＢＭＰ２）。方法将包含ｈＢＭＰ２部分前肽和成熟肽的３′端７３０ｂｐｃＤＮＡ片段，插入原核表达载体ｐｋｐＬ３ａ的多克隆位点，构建成ｐＫｐＬ３ａ／ｈＢＭＰ２重组表达载体，转化大肠杆菌ｐｏｐ２１３６，挑选阳性克隆，经诱导表达后用ＳＤＳＰＡＧＥ鉴定。结果该克隆表达的ｈＢＭＰ２为ＭＷ２７０００，表达量占菌体总蛋白１０％，部分纯化后植入大鼠股部肌肉，组织学观察到肌肉内大量间充质细胞增生及骨与软骨形成。结论说明重组ｈＢＭＰ２具有诱导异位骨与软骨形成的生物学功能。
Objective To explore the way of producing human bone morphogenetic protein 2(hBMP 2) for bone healing by using the gene engeneering techniques. Methods hBMP 2 cDNA fragment, which consists of 3′ end partical propeptide and mature peptide sequence, was inserted into the mutiple cloning site of expression vector pkpL 3a via ligation. The recombinant plasmid pkpL 3a/hBMP 2 was transformed into E.coli pOp2136. By the method of restriction map, the positive expression clone was selected. SDS PAGE analysis showed a new foreign protein band near 27 000 after induction. The yield of induced hBMP 2 accouned for 10% of the total bacterial proteins. Results The partial purified recombinant hBMP 2 was implanted into Wistar rat thigh. After 4 weeks, histological analysis showed that it induced the proliferation of mesenchymal type cells and formation of new cartilage and bone in the implantation area. Conclusion The hBMP 2 produced by gene engeneering techniques has the biologic capacity of ectopic bone formation.
出处
《中华医学杂志》
CAS
CSCD
北大核心
1998年第4期308-310,共3页
National Medical Journal of China
