论文部分内容阅读
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 eukaryotic expression vector pIRES2EGFP-Cadherin-11 and to detect its expression in BMSCs. Methods Cadherin-11 gene was amplified from human Cadherin-11 cDNA by polymerase chain reaction (PCR) and double-digested with Xho I and EcoR I and inserted into vector pIRES2EGFP treated with the same enzymes. The recombinant plasmid Transfected BMSCs, Western blot method was used to detect the expression in cells. Results The results of enzyme digestion and sequencing showed that the amplified Cadherin-11 gene was 2390bp in size and Western blot showed that the Cadherin-11 protein was correctly expressed in BMSCs with a size of about 88kD. Conclusions Cadherin-11 protein is highly expressed in BMSCs and is expected to be used to improve the adhesion between BMSCs and scaffolds.