Salmonella typhimurium X3337lux and applications of salmonella typhimurium X3337lux in living body imaging

A technology of x3337lux and Salmonella typhimurium, applied in the field of microbiology, can solve the problems of high background, difficult quantification, and low specificity of fluorescent labeling in vivo detection, and achieve high specificity and signal-to-noise ratio, stable luminescent quantity, and stable biological The effect of luminous properties

Inactive Publication Date: 2012-10-31
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] 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 X3337lux and its application in live imaging

Method used

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  • Salmonella typhimurium X3337lux and applications of salmonella typhimurium X3337lux in living body imaging
  • Salmonella typhimurium X3337lux and applications of salmonella typhimurium X3337lux in living body imaging
  • Salmonella typhimurium X3337lux and applications of salmonella typhimurium X3337lux in living body imaging

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Experimental program
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Embodiment 1

[0024] Specific construction and identification of strains:

[0025] 1. Material preparation

[0026] 1.1 Plasmids and strains

[0027] 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】

[0028] Salmonella typhimurium ( Salmonella typhimurium ) X3337 was kindly provided by Prof. ROY CURTISS III, School of Biological Sciences, Arizona State University, USA. [HIDENORI MATSUI, CHRISTOPHER M. BACOT, WENDY A. Virulence Plasmid-Borne spvB ​​and spvC Genes Can Replace the 90-Kilobase Plasmid in Conferring Virulence to Salmonella enterica Serovar Typhimurium in Subcutaneously Inoculated Mice[J]. Journal of bacteriology1, 15001 (183): 4652–4658...

Embodiment 2

[0058] Salmonella typhimurium ( Salmonella typhimurium )Χ3337lux 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 NaHCO 3 Gavage, neutr...

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Abstract

The invention discloses a salmonella typhimurium X3337 and applications of the salmonella typhimurium X3337 in living body imaging. The bacterial strain is preserved in depositary authority designated by the state intellectual property office, the preservation data is 13 March, 2012, the depositary authority is China general microbiological culture collection center, and the preservation number is CGMCC No.5894. The salmonella typhimurium X3337 is the bacterial strain after the elimination of virulent plasmids, and can be taken as a reference to be used for researching the functions of the virulent plasmid of salmonella typhimurium. X3337lux is salmonella typhimurium bacterial strain with a stable biological luminescence property. The salmonella typhimurium X3337 complements have the defects that an excitation light source is needed by the detection of bioluminescence bacteria, the specificity of the salmonella typhimurium X3337 in a body is low, the background is high, the quantity is difficult to determine and the like. The salmonella typhimurium X3337 can be widely applied to establishing an in-vitro cell infection model and an in-vivo experiment model. A convenient tool is provided for the intensive study of the pathogenic mechanism of salmonella typhimurium, drug researching and screening and related disease preventing strategy researching and the like.

Description

[0001] technical field [0002] The invention belongs to the technical field of microorganisms, in particular to Salmonella typhimurium X3337lux and its application in live imaging. [0003] Background technique [0004] Salmonella typhimurium ( Salmonella typhimurium ) has a wide range of hosts and can cause a variety of diseases in humans and animals, and it is of great practical significance to carry out research on its pathogenic mechanism. Plasmid is the genetic material outside the bacterial chromosome, which contains many important gene clusters, and is closely related to the symbiosis, metabolism and pathogenic mechanism of bacteria. Salmonella typhimurium X3337 is a strain after eliminating the virulence plasmid, which can be used as a control to study the function of the virulence plasmid of Salmonella typhimurium. [0005] In recent years, in vivo optical imaging technology of living animals has been continuously developed in life science research due to its si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K49/00C12R1/42
Inventor 吴淑燕邱桥成李嫄渊叶颖
Owner SUZHOU UNIV
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