论文部分内容阅读
目的构建Pokemon表达慢病毒载体。方法通过PCR扩增Pokemon cDNA,将Pokemon cDNA连接于GV165载体,经测序确认后,将GV165/Pokemon与pHelper 1.0和pHelper 2.0共转染至293T细胞中,收获病毒,通过Real time PCR测定滴度;将Pokemon表达慢病毒载体侵染293T细胞通过免疫荧光和Western blot检测Pokemon表达慢病毒载体的转染效率和Pokemon的表达能力。结果在感染Pokemon慢病毒载体293T细胞中能检查到Pokemon-GFP融合蛋白的表达。结论成功构建表达Pokemon的慢病毒载体。
Objective To construct Pokemon lentivirus vector. Methods Pokemon cDNA was amplified by PCR. Pokemon cDNA was ligated into GV165 vector. After sequencing, GV165 / Pokemon was co - transfected into 293T cells with pHelper 1.0 and pHelper 2.0. The virus was harvested and titers were determined by Real time PCR. 293T cells were infected with Pokemon expressing lentiviral vector. The transfection efficiency of Pokemon lentiviral vector and the expression of Pokemon were detected by immunofluorescence and Western blot. Results The expression of Pokemon-GFP fusion protein was detected in Pokemon lentivirus 293T cells. Conclusion The lentiviral vector expressing Pokemon was successfully constructed.