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ZIKV(zika virus)-NS1 protein and application of ZIKV-NS1 protein to preparation of ZIKV propagation blocking vaccine

A ZIKV-NS1, 1. ZIKV-NS1 technology, applied to the ZIKV-NS1 protein and its application in the preparation of Zika virus transmission blocking vaccines, can solve problems such as ineffective treatment methods, and achieve the prevention of sudden outbreaks. Outbreak, the effect of reducing the infection rate

Inactive Publication Date: 2018-05-04
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the Zika virus has been discovered for more than 70 years, there is still no specific treatment and effective vaccine that can be applied

Method used

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  • ZIKV(zika virus)-NS1 protein and application of ZIKV-NS1 protein to preparation of ZIKV propagation blocking vaccine
  • ZIKV(zika virus)-NS1 protein and application of ZIKV-NS1 protein to preparation of ZIKV propagation blocking vaccine
  • ZIKV(zika virus)-NS1 protein and application of ZIKV-NS1 protein to preparation of ZIKV propagation blocking vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, the preparation of albumen

[0064] 1. Construction of recombinant plasmids

[0065] The pFastBac-dual vector was used as the starting vector to construct the recombinant plasmid pFastBac-dual-ZIKV-NS1.

[0066] Compared with the pFastBac-dual vector, the difference of the recombinant plasmid pFastBac-dual-ZIKV-NS1 is only that: the double-stranded DNA molecule shown in the sequence 4 of the sequence list is used to replace the double strand shown in the sequence 3 of the sequence list in the starting vector DNA molecule.

[0067] In sequence 4 of the sequence listing, nucleotides 7-1239 are open reading frames encoding the protein shown in sequence 5 of the sequence listing.

[0068] In sequence 4 of the sequence listing, the 7th-9th nucleotide is the initiation codon, the 7th-120th nucleotide encodes the gp67 signal peptide, the 133-162th nucleotide encodes the Myc tag, and the 163-1218th nucleotide The first nucleotide encodes the ZIKV-NS1 protein, a...

Embodiment 2

[0085] Example 2. Application of protein in passive immunization to prevent Zika virus acquisition

[0086] 1. Preparation of mouse antiserum using protein as immunogen

[0087] The experimental animals were 8-week-old Balb / c female mice.

[0088] Myc-ZIKV-NS1-His 6 Proteins are immunogens. The immunogen solution is the Myc-ZIKV-NS1-His prepared in Example 1 6 The protein solution was diluted with PBS buffer.

[0089] Day 1 (initial immunization): test animals were subcutaneously injected with an equal-volume mixture of immunogen solution and Freund's complete adjuvant, and the dose of immunogen given to each test animal was 40 μg (calculated as total protein);

[0090] On the 14th day (the first booster immunization): the test animal was subcutaneously injected with an equal-volume mixture of the immunogen solution and Freund's incomplete adjuvant, and the dose of the immunogen given to each test animal was 40 μg (calculated as total protein);

[0091] Day 28 (second boo...

Embodiment 3、ZI

[0139] Example 3, the application of ZIKV NS1 as Zika transmission blocking vaccine

[0140] The immunogen is Myc-ZIKV-NS1-His 6 protein or OVA protein. The immunogen solution is the Myc-ZIKV-NS1-His prepared in Example 1 6 The protein solution is obtained by diluting the PBS buffer solution or the immunogen solution is obtained by diluting the OVA protein with the PBS buffer solution.

[0141] The experimental animals were 6-week-old AG6 mice.

[0142] 1. Protein active immunization of AG6 mice

[0143] The experimental animals were divided into two groups (the control group used OVA protein as immunogen, and the experimental group used Myc-ZIKV-NS1-His 6 protein as immunogen), 10 mice in each group, the immunization method is as follows:

[0144] Day 1 (initial immunization): test animals were subcutaneously injected with an equal-volume mixture of immunogen solution and Freund's complete adjuvant, and the dose of immunogen given to each test animal was 40 μg (calculate...

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Abstract

The invention discloses ZIKV(zika virus)-NS1 protein and application of the ZIKV-NS1 protein to preparation of ZIKV propagation blocking vaccine. The invention provides protein which is named as the ZIKV-NS1 protein; the ZIKV-NS1 protein is protein formed by amino acid sequences shown as sequence 1 in a sequence table. The invention also provides application of the ZIKV-NS1 protein. The application comprises the following steps (e1) or (e2): (e1) ZIKV vaccine preparation; (e2) ZIKV propagation blocking vaccine preparation. The vaccine provided by the invention can block the propagation of ZIKVthrough mosquitos in nature; the infection rate of mosquitos in natural environment is reduced, so that healthy people groups are protected from being infected by ZIKV; the sudden outbreak of the ZIKV is prevented.

Description

technical field [0001] The present invention relates to ZIKV-NS1 protein and its application in the preparation of Zika virus transmission blocking vaccine. Background technique [0002] Zika virus (ZIKV) is a single-stranded positive-sense RNA virus belonging to the family Flaviviridae, which was first discovered in the jungle of Uganda, Africa. The natural mosquito hosts of Zika virus are Aedes mosquitoes, including Aedes aegypti and Aedes albopictus. Zika virus is spherical and consists of envelope, capsid protein and single-stranded positive-sense RNA. During the evolution of Zika virus, it gradually divided into two lineages—African lineage and Asian lineage. In 2007, Zika virus was only circulating in tropical areas of Africa and Asia until the outbreak in the French Polynesian islands in 2013, followed by a major outbreak in the Americas in 2015. After Zika virus infection, typical clinical symptoms include fever, maculopapular rash, joint and muscle pain. Zika vi...

Claims

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Application Information

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IPC IPC(8): C07K14/18C07K19/00C12N15/40C12N15/62C07K16/10A61K39/12A61K39/42A61P31/14
CPCA61K39/12A61K2039/505C07K14/005C07K16/1081C07K2317/76C07K2319/21C12N2770/24122C12N2770/24134Y02A50/30
Inventor 程功杜森焱
Owner TSINGHUA UNIV
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