Candida albicans attenuated strain with deletion of ARP1 gene and application of candida albicans attenuated strain

A technology for Candida albicans and deletion strains, applied in the field of genetic engineering, can solve the problems of increased mycelium production capacity of Candida albicans, and achieve the effects of reduced toxicity, reduced colonization ability, and weakened proliferation ability

Inactive Publication Date: 2021-03-30
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Nrg1 and Tup1 are more studied, and their deletion will lead to the increase of Candida albicans hyphae production ability (Braun and Johnson, 1997; Murad et al., 2001), and the deletion of ARP1 gene will affect the mycelium production ability of Candida albicans There are no relevant studies and reports on the impact of

Method used

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  • Candida albicans attenuated strain with deletion of ARP1 gene and application of candida albicans attenuated strain
  • Candida albicans attenuated strain with deletion of ARP1 gene and application of candida albicans attenuated strain
  • Candida albicans attenuated strain with deletion of ARP1 gene and application of candida albicans attenuated strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Effect of ARP1 Gene Deletion on Candida Albicans Cell Proliferation

[0037]In order to prove the role of ARP1 deletion in the proliferation of Candida albicans, the present invention carried out the growth curve detection experiment of Candida albicans. Inoculate the wild-type strain of Candida albicans and the ARP1 deletion strain into liquid YPD medium respectively, and after culturing overnight at 30°C, transfer to fresh YPD medium at a ratio of 1 / 100, take samples every two hours to detect OD600, and draw the growth curve , and according to the OD value of the logarithmic phase culture solution, calculate the generation time of the wild-type strain and the ARP1 deletion strain.

[0038] The results are as follows: After Candida albicans was transferred to fresh YPD medium, the OD value of the ARP1 deletion strain at the same time point was significantly smaller than that of the wild-type strain at the same time; the doubling time of the wild-type strain w...

Embodiment 2

[0040] Example 2 The toxicity of Candida albicans with ARP1 deletion is significantly reduced

[0041] In order to prove the influence of ARP1 deletion on the regulation of virulence, the present invention carried out a mouse toxicity experiment. 1.2×10 6 Candida albicans wild strain and ARP1 deletion strain cells were injected into ICR mice, and the survival rate was observed. figure 2 A shows the survival curve of the mice infected with Candida albicans. It was found that the wild-type strain died from the 5th day, and all the mice infected with the wild-type strain died on the 9th day, showing that the average survival time was 6 days; and All mice infected with the ARP1-deficient strain survived up to 15 days, showing no toxicity.

[0042] By detecting kidney sections of dying mice, figure 2 B shows the results of glycogen staining (PAS) of kidney sections of dying mice caused by Candida albicans infection, and the arrows show mycelial cells, showing that the wild-typ...

Embodiment 3

[0043] Example 3 Deletion of the ARP1 gene affects the colonization of Candida albicans in the kidney

[0044] To demonstrate the effect of the ARP1 gene on visceral colonization by Candida albicans. 1×10 6 Candida albicans wild strain and ARP1 deletion strain cells were mixed and injected into 4 six-week-old ICR mice. After 48 hours, kidneys, livers and lungs were collected, ground and homogenized, diluted appropriately and spread on YPD plates and SD-His plates. 30 Count the number of colonies after 2 days of incubation at °C. The result is as image 3 As shown, the proportion of ARP1-deficient strains in liver and lung was 20.8% and 24.3% respectively; while the proportion of ARP1-deficient strains in kidney was only 1.1%. This result indicated that the deletion of the ARP1 gene significantly affected the colonization ability of Candida albicans in the kidney.

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Abstract

The invention provides a candida albicans attenuated strain with deletion of an ARP1 gene and application thereof. The candida albicans attenuated strain with deletion of the ARP1 gene is a candida albicans ARP1 / ARP1 deleted strain without expression of the ARP1 gene, and is prepared through a cytoplasmic division gene ARP1. The Candida albicans attenuated strain lacking the ARP1 gene is used forpreparing or screening Candida albicans therapeutic drugs. The Candida albicans therapeutic drug is a drug which is screened or prepared by taking an ARP1 gene as a target point and enables the ARP1 gene not to be expressed. According to the application of the candida albicans attenuated strain lacking the ARP1 gene in toxicity regulation, the influence of ARP1 gene deletion on the proliferation of candida albicans is detected, thereby detecting the toxicity condition of the candida albicans lacking the ARP1; therefore, the effects of the ARP1 gene-deleted Candida albicans attenuated strain ontoxicity regulation are explored in the process of detecting the influence of the ARP1 gene on visceral colonization and the like. According to the invention, a theoretical basis is provided for treating Candida albicans infection, and a direction is provided for preparing or screening Candida albicans infection treatment drugs.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to an attenuated Candida albicans strain lacking the ARP1 gene and its application. Background technique [0002] Candida albicans (Candida albicans) is one of the most important conditional pathogenic fungi, which can parasitize in the oral cavity, respiratory tract, intestinal tract, and vagina in normal people, and generally do not cause disease or only cause mild skin or mucous membranes, etc. Superficial infection; in people with weakened immunity or immunodeficiency, it can increase its pathogenicity through morphological transformation, etc., causing acute, subacute or chronic invasive fungal infections, resulting in severe severe diseases in the blood system and respiratory system even death. According to statistics, invasive infections caused by Candida albicans account for more than 40% of all Candida infections (Chen M, et al., 2018). At present,...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12Q1/02G01N33/50C12R1/725
CPCC07K14/40C12Q1/02C12Q1/025G01N33/5088G01N2333/40
Inventor 冯金荣冯玉婷姚双燕庄重
Owner NANTONG UNIVERSITY
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