重组人骨形成蛋白-2

摘要： 重组人骨形成蛋白-2(rhBM P-2)具有独特的诱导成骨活性,在修复骨缺损和促进骨愈合方面具有重要作用.细胞学研究表明,rhBM P-2具有良好的促进骨源性细胞及干细胞的成骨功能,即使将rhBM P-2负载于复合材料内依然有较好的促细胞成骨作用.在动物实验中,将rhBM P-2负载于各类复合支架材料上然后植入动物骨缺损模型的骨缺损处均能较好地促进新骨形成.多项临床研究表明,rhBM P-2的临床效果在国内外的临床研究中也均达到预期,因此,将rhBM P-2用于临床骨缺损的修复是可行的.但rhBM P-2临床应用研究最长观察时间只有24个月,其远期效果及安全性仍有待进一步观察.该文对rhBM P-2促骨缺损修复的实验和临床研究的进展进行了综述.

摘要： 目的:观察快速成型(rapid prototyping,RP)技术辅助下制作的个体化假体复合珊瑚羟基磷灰石(coralline hydroxyapatite,CHA)、重组人骨形成蛋白-2(recombinant human bone morphogenetic protein-2,rhBMP-2)修复兔下颌骨缺损的成骨效果.方法:以27只新西兰大白兔为实验对象,随机均分为三组(n=9),全部建立下颌骨连续性缺损模型,并在兔下颌骨缺损区分别植入个体化假体+自体骨(A组)、个体化假体+CHA(B组)及个体化假体+CHA+rhBMP-2(C组).分别于术后4、12、24周三个时间点处死动物取材,进行大体标本观察、骨密度检测、最大抗压力测试,分别比较各组修复骨缺损的能力.结果:术后24周,三组实验兔外形均对称,骨缺损区均有大量新骨形成.术后12、24周,A、C组新骨密度值及最大抗压能力均明显高于B组,差异均有统计学意义(P0.05).结论:RP技术与组织工程技术相结合修复下颌骨缺损,成骨效能肯定,CHA复合rhBMP-2后成骨能力明显增强,为临床应用个体化假体复合人工骨移植修复下颌骨缺损提供可靠的实验依据.

摘要： 目的 探讨重组人骨形成蛋白-2(rhBMP-2)与胶原膜靶向结合后在兔颅骨引导骨再生(GBR)模型中的成骨效应.方法 将20只普通级雌性健康新西兰大白兔随机分成2周组和6周组,每组10只.所有大白兔均制备颅骨GBR模型,在颅顶骨植入4个钛筒,分别盖rhBMP-2/CBD胶原膜(rhBMP-2/CBD胶原膜组)、rhBMP-2胶原膜(rhBMP-2胶原膜组)、胶原膜(胶原膜组)和不盖膜(空白组).分别在2周和6周时处死各对应组的大白兔,取样制作硬组织切片和石蜡切片,染色后进行组织学观察.结果 2周时可见胶原膜阻挡了纤维组织的长入,4组钛筒上层均无新骨生成,其中rhBMP-2/CBD胶原膜组钛筒顶端毛细血管增生量明显较其余3组多.6周时可见rhBMP-2/CBD胶原膜组钛筒上层大量新骨生成,与来源于颅骨骨面的新骨界限明显,而其余3组钛筒顶端未见新骨生成.结论 rhBMP-2与胶原膜靶向结合可形成具有骨诱导性的胶原膜,缓释rhBMP-2使胶原膜下方大量新骨生成,表层成骨可阻止纤维组织的长入和防止植骨床的塌陷,使成骨速度加倍.

摘要： 背景：已有研究证实胶原-羟基磷灰石复合重组人骨形成蛋白2植骨效果较好，然而尚未有文献报道此复合材料与自体骨移植效果的对比评价。n　　目的：检测自体髂骨移植与胶原-羟基磷灰石复合重组人骨形成蛋白2植骨在单侧完全性腭裂大鼠模型中的愈合效果。n　　方法：首先建立32只SD大鼠单侧完全性腭裂模型，随机均分为两组，对照组于裂隙处移植自体髂骨，实验组移植胶原-羟基磷灰石复合重组人骨形成蛋白2生物材料，于移植后的第1，2，3，4周，检测血清中碱性磷酸酶和抗酒石酸酸性磷酸酶活性、新生上腭骨骨密度、成骨细胞特异性标志物骨钙素、骨保护素、核心结合因子及破骨细胞特异性标志物破骨细胞分化因子等基因的表达。n　　结果与结论：随时间的推移，碱性磷酸酶活性逐渐升高，抗酒石酸酸性磷酸酶活性逐渐降低，实验组碱性磷酸酶活性始终高于对照组(P<0.05，P<0.01)，抗酒石酸酸性磷酸酶活性始终低于对照组(P <0.05，P <0.01)；骨密度逐渐增高，实验组始终高于对照组(P <0.01)；骨钙素、骨保护素、核心结合因子基因水平逐渐上升，实验组始终高于对照组；破骨细胞分化因子逐渐降低，实验组始终低于对照组。表明胶原-羟基磷灰石复合重组人骨形成蛋白2复合生物材料植骨方式较自体植骨方式更有优势。%BACKGROUND:It has been demonstrated that the effect of colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft is good, but there is no comparative analysis of this composite graft and autologous bone graft. n OBJECTIVE:To detect the healing effect of autologous bone graft versus colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft in a rat model of unilateral complete cleft palate. n METHODS: Firstly, we established the artificial unilateral complete cleft palate models in 32 Sprague-Dawley rats, and then the established animal models were randomly divided into control group and experimental group. The autologous iliac bone was transplanted into the fissures of control group, and the experimental group received colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft. After that, the activities of serum alkaline phosphatase and tartrate-resistant acid phosphatase, bone mineral densities in the neonatal palate, expressions of osteocalcin, osteoprotegerin, core binding factor, and osteoclast differentiation factor were detected at 1, 2, 3, 4 weeks after treatment. n RESULTS AND CONCLUSION: Over time, the alkaline phosphatase activity increased gradualy, while tartrate-resistant acid phosphatase activity decreased. Compared with the control group, the alkaline phosphatase activity was always higher (P < 0.05,P < 0.01) but the tartrate-resistant acid phosphatase activity was lower in the experimental group (P < 0.05,P < 0.01); the bone mineral density increased in both groups, but it was always higher in the experimental group than the control group (P < 0.01). The expression levels of osteocalcin, osteoprotegerin, and core-binding factor gene gradualy rose in both groups, but they were always higher in the experimental group than the control group; in contrast, the expression of osteoclast differentiation factor was decreased in both groups, and it was lower in the experimental group than the control group. These findings indicate that the colagen/hydroxyapatite/recombinant human bone morphogenetic protein-2 composite graft has more advantages compared with bone autograft.

摘要： 背景：已有研究证实胶原-羟基磷灰石复合重组人骨形成蛋白2植骨效果较好，然而尚未有文献报道此复合材料与自体骨移植效果的对比评价。 目的：检测自体髂骨移植与胶原-羟基磷灰石复合重组人骨形成蛋白2植骨在单侧完全性腭裂大鼠模型中的愈合效果。 方法：首先建立32只SD大鼠单侧完全性腭裂模型，随机均分为两组，对照组于裂隙处移植自体髂骨，实验组移植胶原-羟基磷灰石复合重组人骨形成蛋白2生物材料，于移植后的第1，2，3，4周，检测血清中碱性磷酸酶和抗酒石酸酸性磷酸酶活性、新生上腭骨骨密度、成骨细胞特异性标志物骨钙素、骨保护素、核心结合因子及破骨细胞特异性标志物破骨细胞分化因子等基因的表达。 结果与结论：随时间的推移，碱性磷酸酶活性逐渐升高，抗酒石酸酸性磷酸酶活性逐渐降低，实验组碱性磷酸酶活性始终高于对照组（P〈0.05，P〈0.01），抗酒石酸酸性磷酸酶活性始终低于对照组（P 〈0.05，P 〈0.01）；骨密度逐渐增高，实验组始终高于对照组（P 〈0.01）；骨钙素、骨保护素、核心结合因子基因水平逐渐上升，实验组始终高于对照组；破骨细胞分化因子逐渐降低，实验组始终低于对照组。表明胶原-羟基磷灰石复合重组人骨形成蛋白2复合生物材料植骨方式较自体植骨方式更有优势。

摘要： 目的：通过建立动物骨缺损模型，比较重组人骨形成蛋白-2(rhBMP-2)与珊瑚羟基磷灰石(CHA)复合物、富血小板纤维蛋白(PRF)与珊瑚羟基磷灰石复合物、自体骨与珊瑚羟基磷灰石复合物以及单纯珊瑚羟基磷灰石这四种骨移植材料在骨缺损中的成骨效能。方法：在比格犬双侧胫骨干骺端制备四个相同的骨缺损区，在缺损区分别植入rhBMP-2/CHA、 PRF/CHA、自体骨/CHA及CHA （对照）；3个月后处死动物，行大体标本观察；拍牙科CT，观察各植骨区骨密度情况；制作石蜡切片、 HE染色，比较各植骨区骨组织学特点及新骨形成量。结果：大体标本见四组骨缺损间隙均完全关闭。 X线示自体骨/CHA组和PRF/CHA组骨密度较致密， rhBMP-2/CHA组致密性低于前两者， CHA组未见明显骨致密影。 HE切片见四组新生骨与宿主骨连接紧密，新生骨小梁不规则，粗细不一，排列无序；复合型骨移植材料的新生骨小梁比对照组更密集、粗大，连续性更好；四组植骨区成骨量比较：自体骨/CHA组>PRF/CHA组>rhBMP-2/CHA组>CHA组。结论：复合型骨移植材料成骨效应明显优于单纯珊瑚羟基磷灰石；三种复合型材料中自体骨/CHA成骨效应最好，其次为PRF/CHA， rhBMP-2/CHA最差。%Objective: By establishing bone defects animal model, the osteogenesis effects of four groups bone transplantation materials that they are compound of reco mbinant human bone morphogenetic protein-2 (rhBMP -2), coralline hydroxyapatite (CHA), compound of platelet -rich fibrin and coralline hydroxyapatite, compound of autologous bone and coralline hydroxyapatite and pure coralline hydroxyapatite were compared by repairing bone defects. Methods: Four same bone defects were prepared on each side of tibial metaphyseal of Beagles , rhBMP-2/CHA, PRF/CHA, autologous bone/CHA and pure CHA were respectively implanted into the bone defects. The Beagles were executed after three months, using gross specimen and dental CT to compare four groups bone growth in bone graft areas and bone mineral density; and decalcified paraffin section, HE staining were used to compare histological characteristics and osteogenesis quantity of new bones. Results: Gross specimen: the space between bone defects of four graft areas were completely closed. X-ray showed: the bone densities were more dense of autologous bone/CHA group and PRF/CHA group, the next for rhBMP-2/CHA group, and there is no significant dense bone density shadow in CHA group. HE staining showed that new bones and host bones were connected closely, new bone trabecular was irregular, varied, and the arrangement was disorderly; compared with CHA group, new bone trabeculars on bone graft areas of other three groups were more dense, thick, with better consecu- tiveness; osteogenesis quantity on bone graft areas of four groups were compared as following: autologous bone/CHA group>PRF/CHA group>rhBMP-2/CHA group>CHA group. Conclusion: osteogenesis effects of composite bone graft materials were much better than that of coralline hydroxyapatite alone;autologous bone/CHA was the best in the osteogenesis effects of three kinds of composite bone graft materials, and that of PRF/CHA was followed by, rhBMP-2/CHA was the last.

摘要： 目的：观察rhBMP2（重组人骨形成蛋白-2）与rhVEGF（重组人表皮生长因子）联合用药对体外培养SD大鼠成骨细胞不同成熟阶段的增殖、分化及胎鼠跖骨生长的影响。方法分离新生大鼠颅盖骨的细胞培养获得前成骨细胞（ preosteoblast ， pOB），再诱导分化为成骨细胞（osteoblast，OB），以rhBMP2和低（2μg/L）、中（4μg/L）及高（8μg/L）剂量rhVEGF干预48 h，比较乳酸脱氢酶（ LDH）活性、细胞相对增殖率（ RGR）和碱性磷酸酶（ AKP）活性。另取胎鼠跖骨，以rhBMP2和rhVEGF干预7 d和14 d，比较其纵向骨生长率（ longitudinal bone growth rate ，LBDR）。结果①LDH组间差异无统计学意义。②RGR：pOB的中、高剂量联用组较同剂量rhBMP2组高。 OB的中、高剂量联用组较同剂量rhBMP2及rhVEGF组高。③AKP：pOB的中、高剂量联用组较同剂量rhBMP2及rhVEGF组高。 OB的组间差异无统计学意义。④LBDR；7d联用组较rhBMP2及rhVEGF组高，但14d联用组差异无统计学意义。结论适量联用rhBMP2与rhVEGF对骨再生诱导及骨组织早期生长有协同促进效应。%Objectives To observe the effect of rhBMP2 (recombinate human bone morphogenetic protein-2) combined with rhVEGF ( recombinate human vascular endothelial growth factor ) on the osteoblast proliferation and differentiation and the metatarsal growth in SD rats in vitro.Methods Cells were isolated enzymatically from the cranium of newborn rats .The pre-osteoblasts (pOBs) were cultured and then induced into the osteoblasts (OBs).Then, the cells were cultured with rhBMP-2 (300μg· L-1) and/or rhVEGF in low dose (2 μg· L-1 ), middle dose (4 μg· L-1 ), and high dose (8 μg· L-1 ) for 48h, respectively.The lactate dehydrogenase (LDH) activity, cell relative growth rate (RGR), and alkaline phosphatase (AKP) activity were determined.The metatarsal collected from the SD fetal rats were intervened by rhBMP-2 and/or rhVEGF for 7 days or 14 days.The longitudinal bone growth rate (LBDR) was determined .Results No significant difference of LDH activity among groups was observed .RGR in the middle-and high-dose combined medication groups increased compared with that in the same dose or rhBMP 2 group for pOBs .The results were the same in the middle-and high-dose combined medication groups compared with that in the same dose of rhBMP 2 group and rhVEGF group for OBs.The activity of AKP increased in the middle-and high-dose combined medication groups compared with that in the same dose of rhBMP2 group and rhVEGF group for pOBs , but no significant difference was observed for OBs .LBDR increased significantly in combined medication group of 7-day culturing compared with that in the rhBMP 2 and rhVEGF group , but no significant difference was observed in 14-day culturing .Conclusion The combination of proper dose of rhBMP 2 and rhVEGF can promote osteoblastic generation and bone tissue growth at the early stage in a synergic way .

摘要： Objective To investigate the effect of adding recombinant human bone morphogenetic protein-2 (rhBMP-2 ) on the bone regeneration in sutural supramaxillary distraction osteogenesis (elasticity traction force> 800 g) .Methods 36 healthy hybrid dogs were randomly divided into 5 groups ,control ,400 g ,600 g ,800 g and 800 g+rhBMP-2 (combined) groups ,and killed in postoperative 1-8 weeks for X-ray examination and immunohistology staining .Results Supramaxilla was successfully moved forward in the various traction groups especially in the 600 g group(25.3±5 .2)mm .The traction speed in the 800 g group was fast within 2 weeks and significantly slowed after 4 weeks(12 .1±2 .3)mm ,which was improved after injection of rhBMP-2[(24 .2±3 .4)mm] . Immunohistochemical staining showed that the positive staining of BM P -2 ,BM P-4 and BM P-7 was evident in osteoblast plasma .After injection of rhBMP-2 ,the light density value in BMP-7 was obviously enhanced .The CT examinations showed that the bone density values in jawbone horizontal plate was significantly decreased especially in the 800 g group[(317 .8±185 .5)HU],but after injection of rhBMP-2 ,which was significantly enhanced[(1 158 .0±93 .0)HU ].Conclusion Sutural traction for advancing supramaxilla can extend the bone length at the early stage of microtrauma .RhBMP-2 may play certain role in the bone formation and the regulation of the activity of BMP-7 .%目的 探索上颌骨经缝牵引中(当弹性牵引力大于800 g)加入重组人骨形成蛋白-2(rhBMP-2)对新骨形成的影响.方法 将36只健康杂种犬随机分为5个组,即对照组、400 g组、600 g组、800 g组及800 g+ rhBMP-2组(联合组).术后1~8周处死动物,行X线检测及免疫组织化学染色检测.结果 各牵引组上颌骨成功前徙,以600 g组效果明显[(25.3±5.2)mm].800 g组在2周内牵引速度快,4周后明显减慢[(12.1±2.3)mm],但在注射rhBMP-2后得以改善[(24.2±3.4)mm].BMP-2、BMP-4、BMP-7阳性染色主要定位于成骨细胞细胞质中,BMP-7在注射rhBMP-2后光密度值明显增强.CT检测各牵引组腭骨水平板骨密度值明显减低,以800 g组最为显著[(317.8±185.5)HU],而在注射rhBMP-2后骨密度值明显增强[(1 158.0±93.0)HU].结论 骨缝牵引前移上颌骨能在微创早期延长骨长度;rhBMP-2可以有效促进骨形成并在调节rhBMP-7活性方面起到一定作用.

摘要： 目的：制备重组人骨形成蛋白-2-聚乳酸（rhBMP-2-PL）缓释纳米微球缓释系统，观察其对兔成骨细胞的生物学效应。方法：采用复乳-干燥法制备rhBMP-2纳米微球，观察其一般特性，并模拟体内条件研究BMP-PLA纳米微球的体外缓释特性。采用MTT法检测微球对成骨细胞增殖状况的影响，并与单纯rhBMP-2的作用进行比较。结果：纳米微球表面光滑圆整,球体大小均匀，平均粒径为44nm，rhBMP-2-PLA纳米微球的包封率和载药量分别为(92.14±1.13)%和[(133.81±0.6)×10-3]%。微球缓释系统有生物活性，能显著促进兔成骨细胞的增殖，其效应高于单纯施加rhBMP-2的效应。结论：rhBMP-2-PLA纳米微球缓释系统的作用明显优于单纯rhBMP-2，在骨创伤的治疗中有良好的前景。

摘要： 本发明公开了一种重组人骨形成蛋白-2的表达方法。包括目的基因hBMP-2的合成；用Nde I和BamH I双酶切hBMP-2基因与载体，连接构建重组载体，转化宿主菌，构建hBMP-2基因工程菌；培养hBMP-2基因工程菌，分离、清洗、溶解包涵体得到包涵体溶解液；纯化包涵体；复性；超滤、透析、冻干得到重组人骨形成蛋白-2。本发明工艺简便，成本低，产量高，活性好，复性率可达到60%左右，在产业化生产上具有优势，适于大规模生产。

摘要： 本发明涉及基因工程重组二连体骨形成蛋白的设计构建方法，其特征是用基因工程手段构建“重组人BMP成熟肽二连体”，基本方法是将编码BMP成熟肽的cDNA放在大肠杆菌中表达，使BMP成熟肽在内部正确形成3对双硫健，在BMP成熟肽的链间以肽健相连接而成为BMP二连体以及经在体外纯化复性后折叠成为有活性的BMP；半胱氨酸突变的过程需要加入限制性核酸内切酶位点序列和起始码，并需要PCR的扩增引物；引物根据需要可设计为不同DNA序列；本发明提供了用基因工程的手段使大肠杆菌直接表达类似二聚体结构的有活性的重组二连体骨形成蛋白及其制备方法，以期用廉价的大肠杆菌基因工程系统去大量生产具有良好生物活性的各种BMP。

摘要： 本发明公开了稳定的冠状病毒重组蛋白二聚体及其表达载体，冠状病毒重组蛋白，由冠状病毒S蛋白S‑RBD、冠状病毒N蛋白的CTD区N‑CTD和将二者偶联的连接子构成。本发明一些实例的冠状病毒重组蛋白，可以形成并维持稳定的二聚体结构，避免单体S‑RBD降解，有利于提高冠状病毒重组蛋白的免疫原性，有望用于制备检测试剂原料、疫苗、抗体、预防或治疗性药物。本发明一些实例的冠状病毒重组蛋白二聚体，具有很好的免疫原性。在疫苗开发领域具有广阔的应用前景。本发明一些实例的表达载体，易于表达冠状病毒重组蛋白二聚体且表达量高。

摘要： 本发明公开了一种重组乳铁蛋白二肽、表达载体及其应用。所述重组乳铁蛋白二肽的碱基序列如SEQ ID No.2所示，将包含有该重组乳铁蛋白二肽的表达载体pPICZαA-Lfchimera经限制性内切酶Sac I线性化后经电击转化法转化到毕赤酵母SMD1168中，可成功获得表达。经平板溶菌圈法检测，表达产物具有明显的抑菌活性，进而将其作为绿色添加剂应用于动物饲料中，具有替代抗生素药物的前景。

摘要： 本发明公开了一种重组人骨形成蛋白－2的表达方法。包括目的基因hBMP－2的合成；用Nde I和BamH I双酶切hBMP－2基因与载体，连接构建重组载体，转化宿主菌，构建hBMP－2基因工程菌；培养hBMP－2基因工程菌，分离、清洗、溶解包涵体得到包涵体溶解液；纯化包涵体；复性；超滤、透析、冻干得到重组人骨形成蛋白－2。本发明工艺简便，成本低，产量高，活性好，复性率可达到60％左右，在产业化生产上具有优势，适于大规模生产。

摘要： 本发明属基因工程技术领域。本发明公开了用于修复骨损伤和骨缺损的重组人骨形成蛋白－7成熟肽，其步骤包括：(1)利用反转录聚合酶链式反应获取编码人骨形成蛋白－7成熟肽(羧基端103－139肽)的基因序列；(2)利用基因工程手段获得人骨形成蛋白－7成熟肽的重组表达产物；(3)通过对表达产物的纯化、变性和复性获得具有良好活性的重组人骨形成蛋白－7成熟肽产物；(4)将适量人骨形成蛋白－7成熟肽基因工程产物与植骨材料复合后，能有效地修复多种无法或难以治愈的骨缺损。

摘要： 本发明涉及应用甲醇酵母高效表达人骨形成蛋白技术，尤其涉及根据酵母偏爱密码子对人骨形成蛋白成熟肽序列进行定点突变技术。其特征是用含人BMP4和7cDNA序列的质粒pBluescript KS－BMP4和pBluescript KS－BMP7为模板，PCR扩增得到人BMP4和7成熟肽cDNA序列。利用简化的重叠PCR技术分别对人BMP4和7成熟肽cDNA序列中精氨酸密码子进行定点突变。突变后的序列作为模板克隆到甲醇酵母表达质粒pPIC9K中并转化到甲醇酵母GS115内。通过筛选并获得阳性克隆。构建好的工程菌在BMGY和BMMY培养基中培养，经甲醇诱导后，高效表达重组人骨形成蛋白。本发明方法可高效表达分泌型重组人BMPs。通过定点突变，表达量提高了至少5倍，达到0.2mg/ml以上。重组人BMPs可用于骨科及相关临床治疗。

摘要： 本发明涉及一种能够用于修复骨损伤和骨缺损的重组人骨形成蛋白－2截短突变体及其制备方法,它包括:(1)利用反转录聚合酶链式反应获取编码人骨形成蛋白－2羧基端107肽截短突变体的基因序列;(2)利用基因工程手段获得人骨形成蛋白－2截短突变体的重组表达产物;(3)通过对表达产物的纯化获得具有良好活性的重组人骨形成蛋白－2截短突变体;(4)将适量人骨形成蛋白－2截短突变体基因工程产物与植骨材料复合。经实验证明:本发明能有效地修复多种无法或难以治愈的骨缺损,其活性约为人骨形成蛋白－2完整成熟肽的5倍。

摘要： 本发明公开了用于治疗造血功能障碍和损伤的可溶性重组人骨形成蛋白2,其步骤包括:①利用反转录聚合酶链式反应获取人骨形成蛋白2(rhBMP－2)基因全序列和部分序列;②利用基因工程手段获得了多种人骨形成蛋白2重组表达产物;③通过对表达产物的纯化获得有生物活性的可溶性重组人骨形成蛋白2;④将适量的可溶性重组人骨形成蛋白2肌肉或皮下注射,可对多种造血系统疾病发挥治疗作用。实验证明优于目前临床应用的各种升血因子。

摘要： 本发明涉及基因工程重组二连体骨形成蛋白的设计构建方法，其特征是用基因工程手段构建“重组人BMP成熟肽二连体”，基本方法是将编码BMP成熟肽的cDNA放在大肠杆菌中表达，使BMP成熟肽在内部正确形成3对双硫键，在BMP成熟肽的链间以肽键相连接而成为BMP二连体以及经在体外纯化复性后折叠成为有活性的BMP；半胱氨酸突变的过程需要加入限制性核酸内切酶位点序列和起始码，并需要PCR的扩增引物；引物根据需要可设计为不同DNA序列；本发明提供了用基因工程的手段使大肠杆菌直接表达类似二聚体结构的有活性的重组二连体骨形成蛋白及其制备方法，以期用廉价的大肠杆菌基因工程系统去大量生产具有良好生物活性的各种BMP。