Wang Xiwen, Li Zhiping, Li Bo, Chi Hang, Li Jiakuan, Fan Hongchao, Yao Ruizhi, Li Qianxue, Dong Xiaolin, Chen Man, Qu Han, Wang Yuanyuan, Gao Weicun, Wang Yutian, Sun Yu, Sun Rui, Qian Jun, Xia Zhiping
Key Laboratory of Jinlin Province for Zoonosis Prevention and Control, Military Veterinary Institute, AMMS, Changchun, 130122, China.
Mol Imaging Biol. 2016 Aug;18(4):519-26. doi: 10.1007/s11307-015-0925-6.
The goal of this study was to develop a plasmid-based lux bio-reporter for use to obtain in vivo images of Brucella suis vaccine strain 2 (B.suis S2) infection with high resolution and good definition.
The pBBR-lux (pBBR1MCS-2-lxCDABE) plasmid that carries the luxCDABE operon was introduced into B. suis S2 by electroporation yielding B. suis S2-lux. The spatial and temporal transit of B. suis S2 in mice and guinea pigs was monitored by bioluminescence imaging.
The plasmid pBBR-lux is stable in vivo and does not appear to impact the virulence or growth of bacteria. This sensitive luciferase reporter could represent B. suis S2 survival in real time. B. suis S2 mainly colonized the lungs, liver, spleen, and uterus in mice and guinea pigs as demonstrated by bioluminescence imaging.
The plasmid-based lux bioreporter strategy can be used to obtain high resolution in vivo images of B. suis S2 infection in mice and guinea pigs.
本研究的目标是开发一种基于质粒的发光生物报告基因,用于以高分辨率和良好清晰度获取猪布鲁氏菌疫苗株2(猪布鲁氏菌S2)感染的体内图像。
通过电穿孔将携带luxCDABE操纵子的pBBR-lux(pBBR1MCS-2-lxCDABE)质粒导入猪布鲁氏菌S2,产生猪布鲁氏菌S2-lux。通过生物发光成像监测猪布鲁氏菌S2在小鼠和豚鼠体内的空间和时间转移。
质粒pBBR-lux在体内稳定,似乎不会影响细菌的毒力或生长。这种灵敏的荧光素酶报告基因可以实时反映猪布鲁氏菌S2的存活情况。生物发光成像显示,猪布鲁氏菌S2主要定殖于小鼠和豚鼠的肺、肝、脾和子宫。
基于质粒的发光生物报告基因策略可用于获取小鼠和豚鼠体内猪布鲁氏菌S2感染的高分辨率图像。