Mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging

A kind of technology of a mouse typhoid thabilia and the living imaging, which is used in the field of microorganisms, can solve the problems of low body detection of fluorescent marks, low, difficult to quantitative, high background, etc., to achieve stable biological luminous performance, stable amount of luminous, high -specific and faithful faith The effect of noise ratio

Inactive Publication Date: 2012-10-31
SUZHOU UNIV
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  • Abstract
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Problems solved by technology

[0008] The present invention aims at the defects of low specificity, high background and difficult quantification of fluorescent markers in vivo detection, as well as the shortcomings of previous plasmid-based bioluminescent detection that require additional screening pressure and substrates, and are difficult to quantify, etc., and provides a stable luminescence Salmonella typhimurium UF110lux and its application in live imaging

Method used

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  • Mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging
  • Mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging
  • Mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging

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Experimental program
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Embodiment 1

[0021] Specific construction and identification of strains:

[0022] 1. Material preparation

[0023] 1.1 Plasmids and strains

[0024] Plasmid pBEN276 was kindly donated by Prof. Pierre Germon, Laboratory of Animal Infectious Diseases and Public Health Pathogen Research, French Institute of Natural Resources. 【Kevin Howe, Attila Karsi, Pierre Germon, et.al. Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon [J]. BMC Microbiology 2010, 10:197】

[0025] Salmonella typhimurium ( Salmonella typhimurium ) UF110lux was donated by Prof. ROY CURTISS III, College of Biological Sciences, Arizona State University, USA. [HIDENORI MATSUI, CHRISTOPHER M. BACOT, WENDY A. Virulence Plasmid-Borne spv B and spv C Genes Can Replace the 90-Kilobase Plasmid in Conferring Virulence to Salmonella enterica Serovar Typhimurium in Subcutaneously Inoculated Mice[J]. Journal of bacteriology, 2001, 15(183): 4652–4658.】...

Embodiment 2

[0058] Salmonella typhimurium ( Salmonella typhimurium ) Application of UF110lux in Live Imaging

[0059] 1. Bacterial culture

[0060] Select a single clone from the solid LB agar plate and inoculate it into LB liquid medium, and culture at 37°C, 200rpm for 14-16h to logarithmic growth phase. Collect bacteria by centrifugation at 4000rpm, 4°C, 10min, wash twice with normal saline (4000rpm, 4°C, 10min), resuspend with normal saline, measure OD600 to calculate the bacterial concentration. Gradiently diluted with normal saline for use.

[0061] 2. Mice infection method (intraperitoneal injection or oral feeding infection)

[0062] 2.1 Intraperitoneal injection infection

[0063] Select 8-12 weeks BALB / C or C57BL mice, with 50μL concentration of 10 5 CFU / mL bacterial solution was injected intraperitoneally to infect mice.

[0064] 2.2. Oral feeding infection

[0065] After 8-12 weeks of BALB / C or C57BL mice were fasted for 6 hours, they were treated with 50 μL of 10%wt NaH...

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Abstract

The invention relates to mouse typhus salmonella UF110lux and application of mouse typhus salmonella UF110lux in living body imaging. The strain is conserved in the China General Microbiological Culture Collection Centre specified by the State Intellectual Property Office on March 13, 2012, and the conservation number is CGMCCNo.5893. The mouse typhus salmonella UF110 is a strain obtained by the fact that a spv gene on a virulence plasmid is knockout, and can be used for researching the function of mouse typhus salmonella virulence gene spv as a control. The UF110lux is a strain which is constructed on the basis of the mouse typhus salmonella UF110 and has the stable biological luminescence property. According to the invention, the shortage of a traditional fluorescent mark is overcome, and the invasion, proliferation and diffusion of the mouse typhus salmonella in a living body of mice can be monitored dynamically in real time by using a living body imaging instrument. The invention provides a convenient tool for further study on a pathogenic mechanism of mouse typhus salmonella.

Description

[0001] technical field [0002] The invention belongs to the technical field of microbes, in particular to Salmonella typhimurium UF110lux and its application in live imaging. [0003] Background technique [0004] Salmonella typhimurium ( Salmonella typhimurium ) has a wide range of hosts and a high incidence of infection, and is currently a research hotspot in the fields of food safety, agriculture, animal husbandry and medicine. Salmonella plasmid virulence gene spv (Salmonella plasmid virulence genes) is a highly conserved sequence on the Salmonella virulence plasmid. The encoded product is most closely related to the virulence phenotype of bacteria, can increase the proliferation of Salmonella in extraintestinal tissue cells, and is related to bacterial serum resistance, adhesion and colonization; in addition spv Genes can also affect the intracellular survival of bacteria and the autophagy and apoptosis of host cells. UF110 is on the virulence plasmid of Salmo...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K49/00C12R1/42
Inventor 叶颖吴淑燕李嫄渊黄瑞
Owner SUZHOU UNIV
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