Infuse protein of candida albicans hot shock protein 90 epitope and phage envelop protein , and its uses

A technology of Candida albicans and heat shock protein, applied in the field of DNA recombination, can solve the problems of large side effects, time-consuming, expensive antifungal drugs, etc.

Inactive Publication Date: 2005-01-26
NORTHEAST NORMAL UNIVERSITY
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0003] Systemic Candida albicans infection has no obvious clinical features, preventing clinicians from detecting it at an early stage
At present, the main diagnostic method is to use blood culture, which is not only time-consuming, cumbersome to operate, but also has low sensitivity.
In terms of treatment, antifungal drugs are mainly used for treatment. Antifungal drugs are not only expensive, but also have great side effects, such as easy to cause renal failure, etc.

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0037] 1. Synthesis of epitope gene:

[0038] The epitope gene fragments were performed using a DNA synthesizer from a commercial bio company. The nucleotide sequence of the gene is:

[0039] 5'-CTG AAA GTT ATC AGA AAA-3'

[0040] 2. Construction of recombinant vector

[0041] The plasmid pfd8SHS was used as the original plasmid to construct the recombinant vector.

[0042] 1) Treat the original plasmid pfd8SHS with restriction endonucleases.

[0043] 2) Mix the digested vector with the artificially synthesized exogenous DNA fragment at a molar concentration of 1:3.

[0044] 3) Add 0.5 μL T to 10 μL mixture 4 DNA ligase and 1 μL buffer. Under the condition of 15-20° C. overnight, the recombinant vector (pfd8C) was obtained.

[0045] 3. Preparation of Competent Cells

[0046] TG1 cells were cultured on LB medium at 37°C for 16-20 hours. Pick a single colony and inoculate it in 2ml LB liquid medium, and culture overnight at 37°C. The culture solution was transferred t...

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Abstract

The present invention belongs to DNA recombinant technology field in biological engineering. Using genetic engineering technology, the candida albicans hot shock protein 90 epitope gene fragment (LKVIRK) is inserted into a carrier integrated with silk shape bacterial virus principal coat protein gene to construct a new recombinant carrier. In a host bacterium, the epitope gene product is secreted to the outside of the cell and is assembled in bacterial virus coat protein to form hybrid albumen. 1) The hybrid albumen can be as an antigen for generating immune response to the antibody in the blood serum of the patient infected by system candida albicans. 2) The hybrid albumen generates better humoral immunity and cell immunity in small mouse. 3) The hybrid albumen expresses protection function in the immunity protection and defending experiment for the model mouse infected by the candida albicans. Therefore, the hybrid albumen can be used for preparing the medicine for diagnosing, defending and curing the system candida albicans infection.

Description

technical field [0001] This patent belongs to the technical field of DNA recombination in biological engineering. Background technique [0002] Candida albicans is one of the most common opportunistic pathogens, and its deep (systemic) infection, ie, systemic Candida albicans infection, is life-threatening for patients, and the mortality rate is still high in the case of treatment with antifungal drugs. Candida albicans infection has been the first clinical fungal infection for more than two decades. Research reports have shown that patients with AIDS, cancer, advanced liver disease and a large number of antibiotics are susceptible to Candida albicans, which is one of the common causes of death in patients with weak immune systems. At present, systemic Candida albicans infection is still a threat to human health due to the lack of effective diagnostic reagents and therapeutic drugs. [0003] Systemic Candida albicans infection has no obvious clinical features, preventing c...

Claims

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Application Information

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IPC IPC(8): A61K38/16C07K19/00C12N15/62G01N33/53
Inventor 王丽
Owner NORTHEAST NORMAL UNIVERSITY
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