| 真核表达载体plRES2EGFP—Cadherin-11的构建及其在BMSCs内的表达 |
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| 引用本文： | 柳大烈,王竞鹏,林立新,王晋煌,陈兵,陈伯华. 真核表达载体plRES2EGFP—Cadherin-11的构建及其在BMSCs内的表达[J]. 组织工程与重建外科, 2009, 0(3): 131-133 |
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| 作者姓名： | 柳大烈 王竞鹏 林立新 王晋煌 陈兵 陈伯华 |
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| 作者单位： | [1]南方医科大学附属珠江医院整形美容科,广东省广州市510282 [2]烟台市毓璜顶医院整形美容科,山东省烟台市264000 |
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| 基金项目： | 国家自然科学基金（30570517）. |
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| 摘 要： | 目的构建plRES2EGFP—Cadherin—11真核表达载体，并检测其在BMSCs中的表达情况。方法应用聚合酶链反应技术从人Cadherin-11 cDNA中扩增出Cadherin-11基因后，以内切酶XhoI和EcoRI进行双酶切。将其插入用相同酶处理的载体pIRES2EGFP；将重组质粒转染BMSCs，应用Westernblot方法检测其在细胞中的表达情况。结果酶切和测序结果表明，扩增的Cadherin-11基因序列正确，大小为2390bp；Western blot表明，Cadherin-11蛋白在BMSCs中正确表达，大小约88kD。结论Cadherin—11蛋白能在BMSCs中高度表达，可望用于提高BMSCs与支架材料的粘附性。
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| 关 键 词： | 骨髓间充质干细胞 种子细胞 组织工程 细胞粘附 构建 |
Experimental Study on the Construction of pIRES2EGFP-Cadherin-ll Eukaryotic Vector and the Expression of BMSCs |
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| LIU Dalie,WANG Jingpeng,LIN Lixin,WANG Jinhuang,CHEN Bin,CHEN Bohua. Experimental Study on the Construction of pIRES2EGFP-Cadherin-ll Eukaryotic Vector and the Expression of BMSCs[J]. Journal of Tissue Engineering and Reconstructive Surgery, 2009, 0(3): 131-133 |
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| Authors: | LIU Dalie WANG Jingpeng LIN Lixin WANG Jinhuang CHEN Bin CHEN Bohua |
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| Affiliation: | 1 Department of Plastic Surgery, Zhujiang Hospital, The South Medical University, Guangzhou 510282, China; 2 Department of Plastic and Aesthetic Surgery, Yuhuangding Hospital, Yantai 264000, China ) |
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| Abstract: | Objective To construct pIRES2EGFP-Cadherin-11 eukaryotic vector and detect its expression in BMSCs cells. Methods Cadherin-11 genes were amplified from human cadherin-11（CDH11） cDNA by polymerase chain reaction （PCR） and cloned into pIRES2EGFP. After the recombinant plasmid was transferred into BMSCs, protein expression of the plasmid was examined by Western blot. Results DNA sequencing and enzyme digestion showed that the cloned cadherin-11 gene was correct. It was about 2 390 bp. The expression of cadherin-ll protein in BMSCs were 88 kD. Conclusion Cadherin-11 is a Ca^2＋-dependent homophilic cell adhesion molecule that is expressed mainly in osteoblasts. The data suggested that cadherin-11 protein can express in BMSCs and improve the adhesion between BMSCs and scaffolds. |
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| Keywords: | Bone mesenchymal stem cells Seeding-cells Tissue engineering Cell adhesion Construction |
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