真核表达载体pIRES2EGFP-Cadherin-11的构建及其在BMSCs内的表达  

Experimental Study on the Construction of pIRES2EGFP-Cadherin-11 Eukaryotic Vector and the Expression of BMSCs

作　　者：柳大烈[1] 王竞鹏[1] 林立新[2] 王晋煌[1] 陈兵[1] 陈伯华[1] 

机构地区：[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

出　　处：《组织工程与重建外科杂志》2009年第3期131-133,共3页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金(30570517)

摘　　要：目的构建pIRES2EGFP-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Xho I和EcoR I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Western blot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。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.

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 

分 类 号：Q781[生物学—分子生物学]