构建化学发光-绿色荧光稳定标记的鼠伤寒沙门氏菌

The Construction of Stable Chemiluminescence-Green Fluorescent Marked Salmonella typhimurium

基因改造的鼠伤寒沙门氏菌(Salmonella typhimurium)作为弱毒疫苗活载体被广泛应用于多种病毒、细菌、寄生虫等的免疫研究和生产实践中,甚至在肿瘤治疗方面也有诸多研究。在科学研究中,可视化技术为研究提供十分方便的手段。在鼠伤寒沙门氏菌强毒株ATCC 14028s基因组上插入Lux operon和gfp基因,使其同时具有化学发光和绿色荧光的特征,而这种基因组的改变并未影响其生长和毒力;在荧光显微镜下可观察到其在食管癌细胞TE1和乳腺癌细胞MDA-MB-468的分布情况;用小动物活体成像系统可检测其在小鼠体内的分布情况。该菌株在细胞水平下的绿色荧光信号和在小鼠体内的化学发光信号搜集方便,信号明显,为今后进一步研究鼠伤寒沙门氏菌提供了良好的可视化手段。

Genetically engineered Salmonella typhimurium , as attenuated live vaccine vector, is widely used in immune research and clinical application of a variety of viruses, bacteria, parasites and cancer. Visualization technology is very useful and important in scientific research. In this study, Lux operon and gfp gene were inserted into the genome of S. typhimurium ATCC14028s strain, a virulent strain. Though the growth and virulence weren′t affected, this modification make it had the characteristics of chemiluminescence and green fluorescence. This strain can infect a variety of cancer cells including TE1 and MDA-MB-468 and can be observed under fluorescence microscope. It also can be detected by in vivo imaging system in mice. The green fluorescence signal at the cellular level and chemiluminescence signal in mice of this new strain were convenient to be checked and gathered because of clear signals. This study provided fine visualized means for further researches on Salmonella typhimurium .