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Novel staphylococcus aureus fluorescence labeling system

A staphylococcus, golden yellow technology, applied in bacteria, microorganism-based methods, microorganism determination/inspection, etc., can solve the problems of cumbersome operation process, cumbersome operation, inability to perform real-time observation, etc., and achieve the effect of simplifying operation steps

Inactive Publication Date: 2015-04-22
UNIV OF SCI & TECH OF CHINA
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  • Description
  • Claims
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Problems solved by technology

The traditional intracellular localization needs to be observed through a microscope after cell sectioning and staining. The operation process is cumbersome and cannot be observed in real time.
In bacterial biofilm research, labeling bacteria by acridine orange staining is cumbersome and highly toxic

Method used

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  • Novel staphylococcus aureus fluorescence labeling system
  • Novel staphylococcus aureus fluorescence labeling system
  • Novel staphylococcus aureus fluorescence labeling system

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Embodiment Construction

[0018] The technical solution of the present invention will be described in further detail below. It should be noted that the various embodiments of the present invention can be combined in any way as desired.

[0019] A first aspect of the invention provides shuttle plasmids for stable high-level expression of green and / or red fluorescent proteins.

[0020] In one embodiment, the gene encoding the green and / or red fluorescent protein is placed under the control of a high-expression promoter, so that the green and / or red fluorescent protein can be expressed at a high level.

[0021] In another embodiment, the shuttle plasmid can be efficiently and stably expressed in a variety of Staphylococcus aureus, so that the shuttle plasmid and the corresponding strains become a system or platform for research use. In a preferred embodiment, the shuttle plasmid is pALC1484.

[0022] The second aspect of the present invention provides a bacterial strain comprising the shuttle plasmid of...

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Abstract

By selecting a high-expression promoter, a shuttle plasmid capable of stably expressing green fluorescent protein and red fluorescent protein is constructed, so that various staphylococcus aureus bacterial strains which can be used for stably expressing fluorescent protein at a high level are obtained. The conventional dye labeling is substituted, so that real-time dynamic observation can be realized for observing and dynamically researching biological behaviors of staphylococcus aureus in a biological film formation process, a cell infection process and animal infection process in real time. Fluorescent labels in different colors can be used for effectively analyzing interaction among different bacterial strains, so that technical support is provided for researching the similarities and differences of different staphylococcus aureus in the biological film formation process and a host interaction process.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fluorescently labeled strain of Staphylococcus aureus obtained by using a shuttle plasmid capable of stably expressing green fluorescent protein and / or red fluorescent egg at a high level, its construction method and application. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus, hereinafter referred to as Staphylococcus aureus) is an important pathogen that can produce a variety of exotoxins and enzymes and can cause a variety of infectious diseases. The infection caused ranges from superficial skin infection (such as furuncle, pain, wound suppuration, etc.) to severe invasive disease (such as pneumonia, empyema, meningitis, endocarditis, etc.), and even sepsis, sepsis, Systemic infectious diseases such as toxic shock used to be the main pathogenic bacteria causing death from traumatic infection (1). The reason why Staphylococcus aureus is highly pathogen...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N1/21C12Q1/02C12R1/445
Inventor 文雯张旭赵丽萍
Owner UNIV OF SCI & TECH OF CHINA
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