GFR/DDR2嵌和受体的构建和稳定表达的鉴定

Construction and stable expression of EGFR/DDR2 chimeric receptor

目的 :将一在细胞膜表面不表达或低表达的受体胞外区与盘状结构域受体 2 (DDR2 )的胞内区融合 ,构建一嵌和DDR2受体 ,避免在活化此受体的同时激活与DDR2有相同配体的受体介导的信号通路 ,为探索DDR2介导的胞内信号途径奠定基础 .方法 :采用PCR扩增和限制性酶切的方式 ,将表皮生长因子受体 (EGFR)的胞外区与DDR2的跨膜及胞内区进行融合并稳定转染EGFR低表达的 2 93细胞 ,G4 1 8筛选 ,流式细胞仪检测嵌和受体在膜表面的表达状况 .结果 :成功构建了EGFR/DDR2嵌和表达载体 ;筛选到稳定表达此嵌和受体的 2 93细胞株 .结论 :融合并稳定表达了EGFR/DDR2嵌和受体 .

AIM: To analyze DDR2 signal pathway by avoiding activating a related family member with the same ligand as DDR2. METHODS: The EGFR/DDR2 chimeric receptor was fused by PCR amplification and enzyme digestion and then stably transfected to 293 cell line with relatively low EGFR expression. After screening with G418, positive clones that stably express this chimera on the membrane were identified by flow cytometry (FCM). RESULTS: EGFR/DDR2 chimeric expressing plasmid was successfully constructed and the clone stably expressing the plasmid was screened. CONCLUSION: The EGFR/DDR2 chimeric receptor has been fused and stably expressed.