Optimized expression of dual reporter genes in transient transfection of purified Toxoplasma gondii using different promoters

作者：Hao Yongxin; Yang Jianmin; Li Xuelian; Ding Jun; Zhang Wei; Liu Qun^*

来源：Canadian Journal of Microbiology, 2012, 58(4): 483-489.

DOI：10.1139/W2012-017

• Toxoplasma gondii
• transfection
• luciferase
• fluorescence activated cell sorting
• fluorescent protein
• dual reporter gene

Fluorescent protein and luciferase genes are valuable reporter genes and have been widely used for noninvasive monitoring of gene expression in living tissues and cells. We tested expression of the dual reporter genes in transient transfection of purified Toxoplasma gondii tachyzoites. Two copies of the enhanced yellow fluorescent protein (EYFP) gene were put under the control of 3 representative T. gondii promoters (GRA1, SAG1, and DHFR). Fluorescence from each EYFP reporter was significantly higher than that from a green fluorescent protein (GFP) reporter. The GRA1-EYFP reporter gave the highest fluorescence. Although both fluorescence and luciferase were expressed in the dual reporter system, the luciferase reporter was more efficient than either the EYFP or GFP reporters, and it required fewer parasites to be successfully used.

• 出版日期2012-4
• 单位中国农业大学