基于近红外荧光蛋白的细菌体内成像研究

Establishment of bacteria imaging system based on expression of near-infrared fluorescent protein

目的建立表达iRFP的细菌株并且进行体内和体外成像。方法合成iRFP基因序列,构建pPLBV和pET-3a-iRFP质粒,转化质粒进入细菌,检测细菌激发和发射波长,分析细菌体外和体内荧光分子成像。结果转化含iRFP质粒的细菌的激发波长峰值为690 nm,发射波长峰值为713 nm,细菌可进行体外和体内荧光分子断层成像。结论本研究成功构建了表达iRFP的细菌,体内外成像可快速分析该细菌的数量变化和体内分布。

Objective To establish bacteria imaging system based on expression of near-infrared fluorescent protein （iRFP）. Methods iRFP gene was de novo synthesized. The pPL-BV and pET-3a- iRFP plasmids were constructed and co-transfected into bacteria BL21. The excitation and emission spectra of iRFP-transfected bacteria BL21 were measured. Fluorescence molecular imaging of bacteria in vitro and in vivo were performed. Results The excitation and emission wavelength of iRFP- transfected bacteria was 690 nm and 713 nm, respectively. Fluorescence molecular imaging of iRFP- transfected bacteria was successfully performed in vitro and in vivo. Conclusion A bacterial strain expressing iRFP has been successfully established, which can be used for rapid analysis of bacteria quantity and distribution by in vivo and in vitro imaging.