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Application of candida albicans attenuated strain without HOF1 gene

A technology for Candida albicans and deletion strains, applied in the direction of microorganism-based methods, medical preparations containing active ingredients, bacteria, etc., can solve the problems of loss and weakening of virulence, and achieve the effect of reduced toxicity and reduced viability

Pending Publication Date: 2020-08-07
NANTONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Direct DNA damage repair proteins have also been shown to be associated with virulence: for example, the loss of Candida albicans recombination repair protein Rad52 and non-homologous end junction protein Lig4 can lead to loss of virulence in systemic infections (Bellido A, et al., 2018 ); the histone acetyltransferase Rtt109 of Candida albicans is involved in maintaining genome stability, but its deletion was also found to lead to significantly reduced virulence in cell and mouse infection experiments, and was directly linked to reactive oxygen species (da RosaJ L, et al., 2010)

Method used

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  • Application of candida albicans attenuated strain without HOF1 gene
  • Application of candida albicans attenuated strain without HOF1 gene
  • Application of candida albicans attenuated strain without HOF1 gene

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 HOF1 Effects of gene deletion on the survival of Candida albicans in host cells

[0027] to prove HOF1The function of the missing Candida albicans in interacting with host cells, the present invention carried out the co-cultivation experiment of Candida albicans and macrophages (RAW264.7). Will 5×10 5 Macrophages were seeded into 12-well plates and incubated at 37°C in 5% CO 2 For overnight culture in medium, the wild-type strain of Candida albicans and HOF1 For the deletion strain, unbound Candida albicans was washed away after 2 hours, some of the macrophages were lysed with 0.01% SDS, diluted appropriately and spread on the YPD plate; the rest of the cells continued to be cultured for 6 hours, and the cells were lysed and spread in the same way flat. All the plates were cultured at 30°C for 2 days, and the number of colonies was counted, and the wild bacterial strain and HOF1 Growth of the deletion strain.

[0028] The results are as follows: after...

Embodiment 2

[0029] Example 2 HOF1 Deleted Candida albicans is significantly less virulent

[0030] to prove HOF1 For the effect of deletion on virulence regulation, the present invention has carried out mouse toxicity experiments. 1×10 6 Candida albicans wild strain and HOF1 The cells of the deletion strain were injected into Balb / c mice, and the survival rate was observed-, the results were as follows figure 2 As shown in A, it was found that the wild-type strain began to die on the 5th day, and all mice infected with the wild-type strain died on the 12th day; HOF1 The mice of the deletion strain remained alive until the end of the 21-day observation period, showing a significant reduction in virulence; in addition, the present invention also added high concentrations of (2×10 6 ) wild strain of Candida albicans and HOF1 Inject the cells of the deletion strain into Balb / c mice and observe the survival rate (such as figure 2 Shown in B), after the wild-type strain was injected ...

Embodiment 3

[0032] Example 3 HOF1 Gene deletion affects kidney colonization by Candida albicans

[0033] to prove HOF1 Genetic influence on visceral colonization by Candida albicans. 2×10 6 Candida albicans wild strain and HOF1 The cells of the deletion strain were injected into three five-week-old Balb / c mice, and the right kidney was collected 48 hours later. After being ground and homogenized, it was appropriately diluted and spread on a YPD plate. After culturing at 30°C for 2 days, the number of colonies was counted. The result is as image 3 As shown, in the mice infected with the wild-type strain, the bacterial content in the kidney was 8.1×10 5 / g; while infected HOF1 In mice with the deletion strain, the bacterial content in the kidney was only 1.6×10 3 / g. This result shows that, HOF1 Deletion of the gene significantly affected the ability of Candida albicans to colonize the kidney.

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Abstract

The invention discloses a candida albicans attenuated strain. The candida albicans attenuated strain is a candida albicans hof1 / hof1 deleted strain which is not expressed by an HOF1 gene. The invention further discloses an application of the candida albicans attenuated strain without the HOF1 gene in preparation or screening of candida albicans treatment drugs. The candida albicans treatment drugscan be prepared by making the HOF1 gene not expressed through RNA interference or a gene recombination method, or the HOF1 protein antagonist is prepared, so that the protein expressed by the HOF1 gene does not play a role. The invention further discloses an application of the candida albicans attenuated strain without the HOF1 gene in toxicity regulation and control. The application comprises the following steps: detecting the influence of HOF1 gene deletion on the survival of Candida albicans in host cells, detecting the toxicity condition of the candida albicans lacking HOF1, and detectingthe influence of the HOF1 gene on visceral colonization of Candida albicans. The invention provides a novel medicament and direction for treating candida albicans infection.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to the application of an attenuated Candida albicans strain lacking the HOF1 gene, in particular to an attenuated Candida albicans strain, a deletion HOF1 Application of genetically attenuated Candida albicans strains in the preparation or screening of Candida albicans therapeutic drugs, a deletion HOF1 Application of genetically attenuated strains of Candida albicans in virulence regulation. Background technique [0002] Candida albicans ( Candida albicans ) is the most important human pathogenic fungus, which widely exists in the oral cavity, respiratory tract, intestinal tract and vagina of normal people, generally does not cause disease or only causes mild superficial infections such as skin or mucous membranes; When the mutual restraint of the flora is out of balance, its pathogenic ability is enhanced, which can cause acute, subacute or chronic invasive fungal inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12Q1/04A61K45/00A61P31/10C12R1/725
CPCC07K14/40C12Q1/04A61K45/00A61P31/10
Inventor 冯金荣姚双燕庄重
Owner NANTONG UNIVERSITY
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