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Construction and expression of eukar-yotic expression vector of fusion gene DDR2-Fc
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 3
- DOI:
- 10.3321/j.issn:1000-2790.2007.03.002
- Key Word:
- 盘状结构域受体2;Fc;DS区域;融合表达
Abstract： 目的:构建DDR2/Fc真核表达载体,使其在HEK293细胞中进行表达. 方法:采用PCR技术,分别从大鼠脑组织、含人IgG1 Fc全长互补DNA(cDNA)的质粒中扩增出盘状结构域受体2(DDR2)的DS区域、Fc段,以定向克隆的方法将其串联至真核表达载体pcDNA3.1(-)中,获得重组表达载体DDR2/Fc-pcDNA3.1(-);采用脂质体转染技术转染HEK293细胞; RT-PCR、免疫印迹检测融和蛋白的表达. 结果:DNA测序和限制型酶切鉴定证明两段基因正确克隆至pcDNA3.1(-)的多克隆位点,细胞裂解物中检测到融合蛋白的表达,其分子质量与预期大小一致,该融合蛋白能正确表达于HEK293细胞中. 结论:成功构建了DDR2/Fc融和表达载体,表达了融合蛋白.
- [1]Olaso E,Labrador JP,Wang L,etc.Discoidin domain receptor 2 regulates fibroblast proliferation and migration through the extracellular matrix in association with transcriptional activation of matrix metalloproteinase-2.[J].2002,277(5).
- [2]Xu L,Peng HB,Wu DY,etc.Activation of the discoidin domain receptor 2 induces expression of matrix metalloproteinase 13 associated with Osteoarthritis in mice[J].2005,280(1).
- [3]Wolfgang Vogel.Discoidin domain receptors: structural relations and functional implications[J].1999,13(Suppl 1).
- [4]Pawson T,Vogel W,Alves F,etc.The discoidin domain receptor tyrosine kinases are activated by collagen.[J].1997,1(1).
- [5]K. Migita,K. Eguchi,T. Tsukada,etc.The Role of Protein Kinase in Human Synovial Fibroblast Growth[J].1995,210(3).