Method for recombinant expression of varicella-zoster virus truncation type glycoprotein E and application thereof

A herpes zoster virus, truncated technology, applied in the ELISA detection of anti-varicella-zoster virus-specific immunoglobulin, recombinant expression of varicella-zoster virus truncated glycoprotein E field, can solve a large number of Expensive manpower and material resources

Inactive Publication Date: 2012-06-27
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Due to the use of monoclonal antibody affinity chromatography, extracting a sufficient amount of gpE from the lysate of VZV-infected cells requires a lot of manpower and material resources and is relatively expensive. If the corresponding antigen can be recombined and expressed using gene recombination technology Avoid the disadvantages of the above methods

Method used

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  • Method for recombinant expression of varicella-zoster virus truncation type glycoprotein E and application thereof
  • Method for recombinant expression of varicella-zoster virus truncation type glycoprotein E and application thereof
  • Method for recombinant expression of varicella-zoster virus truncation type glycoprotein E and application thereof

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Embodiment 1

[0037] Embodiment 1, the recombinant expression of varicella-zoster virus (VZV) truncated glycoprotein E (gpE)

[0038] The recombinant expression method of varicella-zoster virus (VZV) truncated glycoprotein E (gpE) (referred to as truncated VZV-gpE) of the present invention comprises the following steps:

[0039] 1. Acquisition of truncated VZV gpE gene

[0040] (1) Design and synthesis of primers for amplifying the VZV gpE gene

[0041] figure 1 Showing the molecular pattern diagram of the wild-type full-length VZV gpE and the recombinant truncated VZV gpE of the present invention, compared with the wild-type full-length VZV gpE, the recombinant truncated VZV gpE of the present invention has removed the transmembrane region and the intracellular region, And a His tag was added at the C-terminal. According to the gpE sequence of the VZV genome published by NCBI (accession number: GenBank AY253715.1), the present invention designs and synthesizes two primers, and introduce...

Embodiment 2

[0086] Embodiment 2, the application of recombinant truncated VZV gpE protein in the ELISA detection of varicella-zoster virus

[0087] Carry out the ELISA detection of varicella-zoster virus with recombinant truncated VZV gpE protein of the present invention, and specific method comprises the following steps:

[0088] 1. Establishment of rgpE indirect ELISA method

[0089] (1) Buffer and reagents for ELISA detection system

[0090] ① Coating buffer: 50mM carbonate buffer, take 1.76g Na 2 CO 3 , 2.86 g NaHCO 3 Add to 900mL deionized water, adjust to pH 9.6, and use a 1L volumetric flask to make up to volume. 0.22μm membrane filtration preservation.

[0091] ② Rinse buffer: Add 8.76g NaCl, 2.42g Tris base, 0.05% Tween (V / V) into 800mL deionized water, adjust the pH to 7.4 with HCl, and make up to volume with a 1L volumetric flask. 0.22μm membrane filtration preservation.

[0092] ③Sample diluent: Add 1% BSA (w / v) to the washing buffer in ②.

[0093] ④Standard product: ant...

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Abstract

The invention discloses a method for the recombinant expression of a varicella-zoster virus truncation type glycoprotein E and application thereof. The method comprises the following steps: guiding a gene of a varicella-zoster virus (VZV) truncation type glycoprotein E (gpE) in which a transmembrane domain and an intracellular domain are removed and an His label is added into a host cell so as to obtain the recombinant varicella-zoster virus truncation type glycoprotein E by expression. The expression method is beneficial to enhancing the expression quantity of a target protein, the downstream purifying operation is simplified, and the large-scale production of the protein can be realized in an easier way; and moreover, the quality between batches is stable. The recombinant protein disclosed by the invention is used as a capturing antigen and can be used for the indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) detection of specific immunoglobulin for resisting a varicella-zoster virus in a plasma specimen, the accuracy of the clinical diagnosis of VZV infection can be enhanced, and the recombinant protein is also used for other fields needing VZV specific immunoglobulin to carry out high-throughput detection.

Description

technical field [0001] The invention belongs to protein recombinant expression method and application thereof, in particular to a method for recombinant expression of varicella-zoster virus truncated glycoprotein E and its ELISA for anti-varicella-zoster virus-specific immunoglobulin application in detection. Background technique [0002] Varicella-Zoster Virus (VZV) belongs to the family Herpesviridae, belongs to the genus Alphavirus, and humans are its only host. Therefore, it is also called human herpesvirus type 3. It can cause two different clinical and pathological symptoms after infection. The initial infection of VZV occurs mostly in adolescence, manifested as generalized vesicular sores (chickenpox). After that, the virus hides in the sensory ganglia. When the carrier is due to low immunity and other reasons, the virus can be reactivated to cause herpes zoster (the virus passes through The nerve spreads, reaches the epidermal cells, forms vesicular rash symptoms al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/38C12N15/85C07K14/04G01N33/569
Inventor 章金刚王卓唐倩吕茂民冯晶晶马玉媛王建国
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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