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目的伯氏疏螺旋体组氨酸激酶Ⅱ(Hk2)和反应调节蛋白Rrp2共同组成一个二元信号系统,控制病原菌侵染哺乳动物所需致病因子的表达,但Hk2的生物学功能有待验证。方法应用同源重组方法得到hk2敲除菌株(hk2-),并利用蜱-鼠动物模型对其生物学功能进行探索。结果 hk2-能够通过人工针刺正常感染小鼠,间接免疫荧光试验显示在吸血的幼蜱和若蜱中肠能够检测到hk2-,定量PCR结果显示hk2敲除并不影响病原菌在若蜱中肠的繁殖,但在蜱叮咬状态下只有50%的小鼠能够被hk2-感染。结论Hk2不影响病原菌从鼠传播到幼蜱及随后在蜱体内的长期寄生,但hk2敲除降低了病原菌在自然条件下(蜱叮咬)感染小鼠的能力,研究结果暗示Hk2在病原菌生活史中的功能是协助病原菌高效地侵染哺乳动物。
Aim To establish a binary signal system that controls the expression of virulence factors required for pathogens to infect mammals. However, the biological function of Hk2 remains to be verified. Methods Hk2 knockout strain (hk2-) was obtained by homologous recombination method and its biological function was explored by tick-mouse animal model. Results hk2- was able to infect mice normally by artificial acupuncture. Indirect immunofluorescence assay showed that hk2- could be detected in the bloodsucking young ticks and rodents. Quantitative PCR results showed that hk2 knockdown did not affect pathogen in rodents , But only 50% of the mice in the tick bite status were able to be infected with hk2-. Conclusions Hk2 does not affect the transmission of pathogens from mice to young ticks and then to long-term parasites in ticks, but hk2 knockdown reduces the ability of pathogens to infect mice under natural conditions (tick bites), suggesting that Hk2 is involved in the life history of pathogens The function is to help pathogens efficiently infect mammals.