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| 人骨保护素(OPG)重组腺病毒的制备及其生物活性研究 | |
| 引用本文： | 刘继中,纪宗玲,胡蕴玉,陈苏民,朱帮福,杨彤涛.人骨保护素(OPG)重组腺病毒的制备及其生物活性研究[J].生物工程学报,2003,19(1):35-40. |
| 作者姓名： | 刘继中 纪宗玲 胡蕴玉 陈苏民 朱帮福 杨彤涛 |
| 作者单位： | 1. 第四军医大学西京医院全军骨科研究所,西安710032 2. 第四军医大学生物化学与分子生物学教研室,西安710032 3. 第四军医大学唐都医院骨科,西安710032 |
| 摘 要： | 采用RT-PCR法得到人OPG的编码区cDNA，克隆至穿梭质粒pShuttle，构建重组有OPG编码区cDNA的腺病毒DNA，经Pac I 酶切线性化，在脂质体介导下转染HEK293细胞，制备重组腺病毒并测定病毒滴度约为5×106～1.5×107 pfu/mL。体外感染小鼠成肌细胞C2C12，Western blot及ELISA检测证实有OPG蛋白的表达，并可在细胞培养上清中持续表达6周。感染OPG重组腺病毒的C2C12细胞生长状态良好、细胞周期无明显变化。将重组腺病毒加入体外培养的小鼠骨髓细胞的培养基中，诱导形成的破骨细胞数量及在象牙片上形成的吸收陷窝的数量显著减少（P<0.01）。
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| 关 键 词： | 骨保护素， 重组腺病毒， 破骨细胞 |
| 文章编号： | 1000-3061(2003)01-0035-06 |
| 修稿时间： | 2001年7月24日 |
Construction and Biological Activity Study of Human Osteoprotegerin Expressing Adenoviral System |
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| LIU Ji Zhong,JI Zong Ling,HU Yun Yu,CHEN Su Min,ZHU Bang Fu,YANG Tong Tao.Construction and Biological Activity Study of Human Osteoprotegerin Expressing Adenoviral System[J].Chinese Journal of Biotechnology,2003,19(1):35-40. | |
| Authors: | LIU Ji Zhong JI Zong Ling HU Yun Yu CHEN Su Min ZHU Bang Fu YANG Tong Tao |
| Institution: | The Fourth Military Medical University, Department of Orthopedics, Xijing Hospital, China. |
| Abstract: | Using the isolated total RNA from osteosacoma cell line MG63,the cDNA encoding human OPG was amplified by RT PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices |
| Keywords: | osteoprotegerin recombinant adenovirus osteoclast |
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