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Candida albicans secreted aspartic proteinase (Sap2) coding epitope sequence-synthesizing and bacteriophage shell protein-bonding hybrid protein and use thereof

A technology of Candida albicans and hybrid protein, applied in the field of DNA recombination, can solve problems such as high price

Inactive Publication Date: 2011-01-26
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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 nephrotoxicity, etc.

Method used

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  • Candida albicans secreted aspartic proteinase (Sap2) coding epitope sequence-synthesizing and bacteriophage shell protein-bonding hybrid protein and use thereof
  • Candida albicans secreted aspartic proteinase (Sap2) coding epitope sequence-synthesizing and bacteriophage shell protein-bonding hybrid protein and use thereof
  • Candida albicans secreted aspartic proteinase (Sap2) coding epitope sequence-synthesizing and bacteriophage shell protein-bonding hybrid protein and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0044] 1. Synthesis of epitope gene:

[0045] The epitope gene fragments were performed using a DNA synthesizer from a commercial bio company. The nucleotide sequence of the gene is:

[0046] 5’-tct ttg gct caa gtc aaa tat act tct gct tcc agt att-3’

[0047] 2. Construction of recombinant vector

[0048] The plasmid pfd88 was used as the original plasmid to construct the recombinant vector.

[0049] 1) Treat the original plasmid pfd88 with restriction endonucleases.

[0050] 2) Mix the digested vector with the artificially synthesized exogenous DNA fragment at a molar concentration of 1:3.

[0051] 3) Add 1 μL T to 10 μL mixture 4 DNA ligase and 1 μL buffer. Under the condition of 16° C. overnight, the recombinant vector (pfd8V long) was obtained.

[0052] 3. Preparation of Competent Cells

[0053]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 solu...

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Abstract

The invention belongs to the technical field of DNA recombination in bioengineering and particularly relates to a hybrid protein and use thereof. The hybrid protein can synthesize an epitope sequence for coding candida albicans secreted aspartic proteinase (Sap2) and can be bonded with the carrier of a phagemid. According to the analysis on the sequence of the bonded product, the coded protein sequence has 88 amino acids, and the molecular weight of the protein is 6.5kDa. The bonded carrier is used for expressing the hybrid protein in cell of Escherichia coli. The protein has high application values in detection and prevention of systemic Candida albicans infection.

Description

technical field [0001] The invention belongs to the technical field of DNA recombination in bioengineering, and specifically relates to a hybrid protein and its application. Background technique [0002] Candida albicans is a normal flora that inhabits the skin and mucous membranes of healthy hosts. When the body's immunity is weakened or the balance of normal flora is destroyed, it will invade deep tissues and cause systemic Candida albicans infection. It is a yeast-like fungus that can exist in various forms such as spores, yeast and mycelium under different conditions, and the antigens expressed in different forms are also different. [0003] With the use of tumor radiotherapy, chemotherapy and a large number of broad-spectrum antibiotics, the incidence of systemic Candida albicans infection has been increasing. The lack of obvious clinical symptoms of Candida albicans infection makes early diagnosis difficult. Clinically, blood culture method is often used. This metho...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62G01N33/569A61K39/00A61P31/10
Inventor 王丽苏全平
Owner NORTHEAST NORMAL UNIVERSITY