中文摘要：

目的：构建胆碱乙酰基转移酶（ChAT）过表达的慢病毒载体,转染小鼠腹腔巨噬细胞系RAW264.7,上调细胞中ChAT表达.方法：利用PCR方法从小鼠cDNA文库中调取ChAT基因,克隆至pGC-FU慢病毒载体上,经PCR和测序鉴定.用pGC-FU-ChAT质粒转染RAW264.7后,荧光显微镜下观察绿色荧光蛋白（GFP）表达,real-time PCR检测RAW264.7细胞中ChAT基因表达丰度.结果：融合表达GFP的慢病毒载体pGC-FU-ChAT感染RAW264.7细胞的效率可达80%以上,感染细胞ChAT基因表达大幅上调.结论：成功构建了携带ChAT基因的慢病毒载体,能有效感染RAW264.7细胞,细胞中ChAT基因过表达.

英文摘要：

Objective： To construct a lentivirus vector expressing choline acetyltransferase （CHAT）, then to upregulate ChAT over-expression in macrophages RAW264.7 of mice by transfecting the constructed virus. Methods.. ChAT sequence was amplified from mice cDNA, which was inserted into pGC-FU, and identified by PCR and DNA sequencing. The pGC-FU-ChAT lentivirus was transfected into mice RAW264.7 cells. ChAT expression was detected by observing GFP expression with fluorescent microscope and by analyzing ChAT expression level with real-time PCR method. Results： The rate of RAW264. 7 carrying pCA2-FU- ChAT infused with GFP was over 80%, and the infected cells strongly expressed ChAT gene. Conclusions： pGC-FU- ChAT lentivirus vector is successfully constructed, and it can effectively transfect RAW 264. 7 cells. The infected cells over-express ChAT gene.