小鼠甲胎蛋白与结核杆菌热休克蛋白70基因融合载体的构建及体外表达

目的 构建小鼠甲胎蛋白(α-Fetoprotein，AFP)与结核杆菌热休克蛋白70(Mycobacterium tuberculosis heat shock protein 70，Mt．HSP70)基因融合载体．并研究其在真核细胞中的表达情况。方法 以pcDNA3.1为基本单位构建AFP及HSP70的融合表达载体，融合基因以G-S-G-G-S连接子连接；用脂质体将系列载体导入COS-7细胞．48h后以免疫化学方法检测其表达。结果 构建的AFP和HSP70的单独及融合表达载体导入COS-7细胞48h后经RT-PCR检测可扩增出相应片段，细胞免疫化学染色为阳性。结论 AFP和HSP70的单独及融合表达载体构建成功，并能在真核细胞中表达。

Construction and expression of chimeric AFP-HSP70 plasmid in COS-7 cells

Objective To construct chimeric vector of AFP HSP70 and investigate transientexpression of vectors in eukaryotic cells.Methods Vectors were constructed by routine molecular technique. Fusion genes were linked with G S G G S linker; Vectors were transfected into COS 7 cells by Lipofectamine 2000.MRNAs were detected by RT PCR. Proteins were assayed by immunocytochemistry.Results Proteins expressed in COS 7 cells were confirmed by RT PCR and immunocytochemistry 48 hours later.Conclusion AFP HSP70 chimeric vectors are constructed successfully and can express in eukaryotic cells.