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Schistosoma japonicum heat shock protein (SjHSP90) recombinant protein and application thereof to schistosomiasis diagnosis and treatment effect evaluation

A recombinant protein, schistosomiasis technology, applied in the field of immunology, can solve the problem of no SjHSP90 recombinant protein, and achieve the effect of a broad market space

Active Publication Date: 2014-04-16
JIANGSU INST OF PARASITIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no research reports on the application of the SjHSP90 recombinant protein of the present invention to the diagnosis and efficacy assessment of schistosomiasis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Amplification of Schistosoma japonicum Heat Shock Protein SjHSP90 Gene

[0024] Primers were designed according to the gene sequence of Schistosoma japonicum heat shock protein SjHSP90 and synthesized by Shanghai Sangon Bioengineering Co., Ltd.

[0025] Primer P1: 5'-GCCATATGCC TGAACCAATG TCTACC-3', introduce restriction site Nde ;

[0026] Primer P2: 5'-GCCTCGAGCT AGTCCACTTC CTCCATTCC-3', introduce restriction site Xho , the full-length SjHSP90 gene (2160bp) was amplified by PCR.

Embodiment 2

[0027] Example 2: Expression and purification of SjHSP90

[0028] The successfully amplified SjHSP90 gene was directional cloned into the pET28a(+) vector by double enzyme digestion, and confirmed by enzyme digestion and sequencing to obtain the recombinant plasmid pET28a-SjHSP90. The recombinant plasmid was introduced into Escherichia coli BL21 (DE3) competent cells, and the expression of the target protein was induced by IPTG. 12% SDS-PAGE electrophoresis showed that the molecular weight of the fusion protein was about 82kDa, which was consistent with the predicted molecular weight. The bacterial supernatant was further purified by metal chelate column affinity chromatography, and the purity of SjHSP90 recombinant protein reached over 95%, and the yield could reach 200mg / L.

Embodiment 3

[0029] Embodiment 3: the establishment of the animal model of schistosomiasis infection and treatment

[0030] 1) Take healthy New Zealand white rabbits and infect 200 cercariae of Schistosoma japonicum in the abdomen. After 45 days of infection, schistosomiasis eggs are detected in feces to confirm the infection, and then treated with praziquantel at 150 mg / kg body weight, and re-treated with the same dose after 1 week Once; before infection, after infection (before treatment) and after treatment, blood was collected from the ear vein every 1w until 16w after treatment;

[0031] 2) Take Balb / c mice and infect 20 cercariae of Schistosoma japonicum in the abdomen. At the 5th week of infection, schistosomiasis eggs are detected in feces to confirm the infection, and treated with praziquantel at 300 mg / kg body weight, and the same dose after 1 week Retreatment again; before infection, after infection (before treatment) and after treatment, blood was collected from the tail vein e...

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Abstract

The invention relates to a schistosoma japonicum heat shock protein (SjHSP90) recombinant protein and application thereof to schistosomiasis diagnosis and treatment effect evaluation and belongs to the field of immunology. The invention provides the preparation of the SjHSP90 recombinant protein, wherein the amino acid sequence of the SjHSP90 recombinant protein is shown as SEQIDNO.1. The SjHSP90 recombinant protein is further applied to schistosomiasis diagnosis and treatment effect evaluation. According to the invention, the SjHSP90 recombinant protein is applied to the schistosomiasis diagnosis for the first time; the SjHSP90 recombinant protein can be used for not only the schistosomiasis diagnosis and but also the treatment effect evaluation, so that the SjHSP90 recombinant protein has broad market space and good social and economic benefits.

Description

technical field [0001] The invention relates to a heat shock protein SjHSP90 recombinant protein of schistosomiasis japonicum and its application in diagnosis and curative effect assessment of schistosomiasis, belonging to the field of immunology. Background technique [0002] Schistosomiasis is an infectious disease that seriously endangers human health. At present, schistosomiasis is endemic in 74 countries around the world, 779 million people are threatened by schistosomiasis, and more than 200 million people are infected with schistosomiasis. my country is the country with the most serious prevalence of schistosomiasis. It is mainly prevalent in twelve provinces, municipalities, and autonomous regions along the Yangtze River and its south. By 2012, the number of schistosomiasis was still as high as 240,000. Diagnosis is central to the field of schistosomiasis control. Commonly used schistosomiasis etiological diagnosis methods, due to their low sensitivity, high missed ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/70G01N33/68
CPCC07K14/43559C12N15/70
Inventor 王晓婷戴洋戴建荣邢云天曲国立唐建霞赵松朱荫昌
Owner JIANGSU INST OF PARASITIC DISEASES
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