中文摘要：

目的 构建含粉尘螨变应原第2组分（Der f 2）基因真核表达载体,并观察其在小鼠骨髓间充质干细胞（BMSCs）中的表达。方法 提取粉尘螨总RNA,采用反转录聚合酶链式反应（RT-PCR）法扩增Der f 2基因,测序鉴定后将Der f 2基因克隆入pcDNA3.0质粒中,构建真核表达载体pcDNA3.0-Der f 2。将小鼠BMSCs分3组进行实验,包括pcDNA3.0-Der f2转染组、pcDNA3.0空质粒转染组及未转染组,分别利用RT-PCR、蛋白免疫印迹法（Western blot）检测Der f 2基因及蛋白在3组BMSCs中的表达。结果 扩增的Der f 2基因序列与Gen Bank的参考序列完全一致,Der f 2基因正确克隆至真核表达载体pcDNA3.0中。转染BMSCs 48 h后,RT-PCR、Western blot检测结果显示,未转染组和pcDNA3.0空质粒转染组未检测到Der f 2基因表达,pcDNA3.0-Der f 2转染组检测到特异性条带,且大小与预期相符。结论 pcDNA3.0-Der f 2真核表达载体构建成功,Der f 2的mRNA和蛋白能在小鼠BMSCs中正确表达。

英文摘要：

Objective To construct eukaryotic expression vector for group 2 allergen of Dermatophagoides farina（Der f 2）,and observe its expression in mice bone marrow mesenchymal stem cells （BMSCs）.Methods Total RNA was extracted from Dermatophagoides farina,then Der f 2 gene was amplified by reverse transcription polymerase chain reaction（RT-PCR）.Der f 2 gene was cloned into pcDNA3.0 plasmid after sequencing,then eukaryotic expression vector for pcDNA3.0-Der f 2 was constructed.BMSCs of mice were divided into pcDNA3.0-Der f 2 transfection group,pcDNA3.0 transfection group,or the control group.Gene and protein expression of Der f 2 in three groups were separately detected by RT-PCR and Western blot .Results Amplified Der f 2 gene sequences were coincident with the reference sequences published in GenBank ,and Der f 2 gene was correctly cloned into eukaryotic expression vector pcDNA 3.0.After 48 hours of transfection ,the results of RT-PCR and Western blot showed that the expression of Der f 2 gene was not detected in the pcDNA 3.0 transfection group or the control group ,and specific strips were detected only in pcDNA 3.0-Der f 2 transfection group ,and the sizes of strips were coincident with the expectant sizes .Conclusion Eukaryotic expression vector of pcDNA 3.0-Der f 2 can be successfully constructed , and gene and protein of Der f 2 can express correctly in BMSCs of mice .