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人骨保护素编码区基因的克隆及其真核表达载体的构建 被引量：1

Cloning of human osteoprotegerin and construction of mammalian expression vector pcDNA3.1-OPG-IRES-EGFP
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摘要 目的克隆人骨保护素(OPG)编码区基因并构建其真核表达载体。方法以人骨肉瘤细胞系MG63总RNA为模板,采用RT-PCR方法获取OPG编码区基因核苷酸序列,应用定向克隆技术,构建真核表达载体pcDNA3.1-OPG-IRES-EGFP。结果获取的OPG编码区全长核苷酸序列与文献报道的完全一致,经PCR和多酶切鉴定证实构建的表达载体正确。结论获得了人骨保护素(OPG)编码区基因,并成功构建了其真核表达载体。 Objective To clone the encoding gene of human osteoprotegerin（hOPG）and construct the eukaryotic expression vector pcDNA3.1-OPG-IRES-EGFP.Methods Using the isolated total RNA from osteosacoma cell line MG63 as a template,the cDNA encoding hOPG was amplified by reverse transcription-polymerase chain reaction method.Further,the PCR product was cloned into pGEM-T Easy and sequenced.Finally,Recombinant vector pcDNA3.1-OPG-IRES-EGFP was constructed by directionally cloning the target gene fragments into eukaryotic expression vector pcDNA3.1（＋）.Results The OPG cDNA was identical with related literatures,and the recombinant vector pcDNA3.1-OPG-IRES-EGFP was verified by extensive restriction digests and PCR.Conclusion The OPG cDNA is obtained and the recombinant plasmid is constructed successfully.
作者 王晋东 王岩 周勇刚 王冉东 俞广 白金柱 黄鹏 张国强 柴伟
机构地区 中国人民解放军总医院骨科医院
出处 《山西医科大学学报》 CAS 2007年第8期673-676,共4页 Journal of Shanxi Medical University
关键词 骨保护素 基因克隆 载体构建 osteoprotegerin gene cloning vector construction
分类号 Q782 [生物学—分子生物学]
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参考文献9

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同被引文献7

  • 1陈丽萍,杨富生,吴礼安.骨保护素的有关研究[J].牙体牙髓牙周病学杂志,2007,17(6):361-363. 被引量：7
  • 2Simonet WS, Lacey DL, Dunstan CR, et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density[J]. Cell, 1997, 89(2): 309-319.
  • 3Thomas RJ, Guise TA, Yin JJ, et al. Breast cancer cells interact with osteoblasts to support osteoclast formation [J ]. Endocrinology, 1999, 140(10): 4451-4458.
  • 4Kozlow W, Guise TA. Breast cancer metastasis to bone: mechanisms of osteolysis and implications for therapy [J]. J Mammary Gland Biol Neoplasia, 2005, 10(2): 169-180.
  • 5Guise TA, Yin J J, Thomas R J, et al. Parathyroid hormonerelated protein (PTHrP)-(1-139) isoform is efficiently secreted in vitro and enhances breast cancer metastasis to bone in vivo[J].Bone, 2002, 30(5): 670-676.
  • 6Holen I, Cross SS, Neville-Webbe HL, et al. Osteoprotegerin (OPG) expression by breast cancer cells in vitro and breast tumours in vivo--a role in turnout cell survival [J]. Breast Cancer Res Treat, 2005, 92(3): 207-215.
  • 7Corey E, Brown LG, Kiefer JA, et al. Osteoprotegerin in prostate cancer bone metastasis[J]. Cancer Res, 2005, 65 (5): 1710-1718.

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山西医科大学学报
2007年 第8期

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