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Schistosoma japonicum zinc finger protein coding gene SjZFP1 and application thereof

A technology encoding gene, schistosomiasis zinc, applied in application, gene therapy, genetic engineering and other directions, can solve problems such as difficult to achieve on-site control

Inactive Publication Date: 2011-01-19
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some promising candidate vaccine molecules have been found, such as Sj23 (Shi Fuhui, Ye Ping, Lin Jiaojiao, etc., Research on Genetic Engineering Vaccine of Schistosoma japonicum (Mainland China Strain), Chinese Journal of Schistosomiasis Control, 1998, 10 (S1) : 54-59) and Sj28 ((Shi Fuhui, Ye Ping, Lin Jiaojiao, etc., Research on Genetic Engineering Vaccine of Schistosoma japonicum (Chinese Mainland Strain), Chinese Journal of Schistosomiasis Control, 1998, 10(S1): 54-59), However, the protection induced by their use alone is still difficult to meet the needs of on-site control, and it is necessary to continue to screen new candidate vaccine molecules

Method used

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  • Schistosoma japonicum zinc finger protein coding gene SjZFP1 and application thereof
  • Schistosoma japonicum zinc finger protein coding gene SjZFP1 and application thereof
  • Schistosoma japonicum zinc finger protein coding gene SjZFP1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Screening of target genes related to Schistosoma japonicum resistance in Oriental vole

[0020] 1. Materials

[0021] 1.1 oriental vole serum

[0022] Dongting Lake strain orientalis vole was provided by Shanghai Xipuer-Bikay Experimental Animal Co., Ltd. Pick the eyeballs of vole orientalis for blood collection. After taking the whole blood, place it at room temperature. After the blood coagulates, place it in the refrigerator at 4°C for 1 hour, centrifuge at 3000g×20min, take the supernatant, and measure the protein concentration by ultraviolet absorption method.

[0023] 1.2 Phage display cDNA library of Schistosoma japonicum adults

[0024] Schistosoma japonicum adult phage-displayed cDNA library was constructed by the applicant. For the construction method, see Sun Yi, Jia Renchu, Liu Jinming, Yuan Chunxiu, etc., Construction of Schistosoma japonicum adult phage-displayed cDNA library, Chinese Journal of Parasitology and Parasitic Diseases, 2007 , 25(...

Embodiment 2

[0044] Example 2: Observation of the immune effect of phage-displayed Schistosoma japonicum zinc finger protein antigen on mouse schistosomiasis

[0045] 1. Materials

[0046] Phage clones and host bacteria: Select the No. 1-8 clones obtained in Example 1 by screening the phage display library of Schistosoma japonicum adult worms with the serum of orientalis vole, and the EST sequence displayed by No. 1 clone is 74~74~ of the coding gene SjZFP1 of Schistosoma japonicum zinc finger protein. 640bp nucleotide sequence.

[0047] Reagents: HRP-rabbit anti-mouse IgG is the product of TaKaRa Company, the soluble TMB substrate chromogenic solution is the product of Tiangen Biological Company, and the 206 adjuvant (ESSAI ISA 206 CELL 642105) is provided by the French company Sepic. Other reagents are imported or domestic analytical reagents.

[0048] Experimental animals and cercariae of Schistosoma japonicum: Male BALB / c mice (about 18-22 g) aged 4-6 weeks were purchased from Shangh...

Embodiment 3

[0075] Example 3: Cloning and expression of the Schistosoma japonicum zinc finger protein gene SjZFP1 and observation of the immune effect of the recombinant protein

[0076] 1. Materials

[0077] Reagents: Trizol Reagent, Magnetic Beads mRNA Extraction Kit, SuperScript TM II Reverse Transcriptase is a product of Invitrogen. ToPure TM Gel DNA Extraction Kit is a GeneTech product. GenExtract TM Plasmid DNA mini-rapid extraction kit was purchased from Daopu Biotechnology (Beijing) Co., Ltd. Isopropanol, Ethanol, Chloroform, NaCl, KCl, Na 2 HPO 4 、KH 2 PO 4 It is a product of Shanghai Sinopharm Chemical Reagent Co., Ltd. Ampicillin (Amp) is a product of Shanghai Sangon Bioengineering Co., Ltd. TaKaRa Taq TM , TaKaRa Ex Taq, Agarose Regular, and DL2000 were purchased from Treasure Bioengineering (Dalian) Co., Ltd. Tris is a product of Shanghai Weibian Technology and Trade Co., Ltd. Tryptone and Yeast Extract are products of British OXOID company. Agar powder was pu...

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Abstract

The invention belongs to the field of biogen. About 510000 people suffer from Schistosoma japonicum in China, and 65 thousands of people are threatened. The development of an effective vaccine for resisting the Schistosoma japonicum is an important strategy. The invention aims to provide a candidate vaccine molecule for resisting Schistosoma japonicum and an application thereof in preparing recombinant vaccine for resisting Schistosoma japonicum. The invention provides a Schistosoma japonicum zinc finger protein coding gene SjZFP1 as well as a nucleotide sequence thereof and a corresponding coding amino acid sequence thereof disclosed in SEQ IDNO:1. The invention also provides an application of the SjZFP1 gene in preparing the recombinant vaccine for resisting Schistosoma japonicum; the candidate antigens of the recombinant vaccine for resisting the Schistosoma japonicum prepared by utilizing the gene uses an antigen displayed by T7 bacteriophage and a recombined SjZFP1 gene antigen which both have higher immunoprophylaxis effect.

Description

technical field [0001] The invention belongs to the field of biological genes, and in particular relates to the application of the Schistosoma japonicum imago zinc finger protein coding gene SjZFP1 and its product in the preparation of anti-schistosomiasis vaccines. Background technique [0002] Schistosomiasis (Schistosomisis japonica) is a serious zoonotic parasitic disease, 650 million people are threatened worldwide, and about 193 million people are infected with schistosomiasis. Among them, schistosomiasis is mainly prevalent in China and some other Asian countries. In China alone, about 840,000 people are infected, and 65 million people are threatened. Schistosoma-susceptible animals include humans, cattle, sheep and other livestock and wild animals, involving more than 40 kinds of mammals. Livestock infection with schistosomiasis not only causes heavy losses to animal husbandry production, but more importantly, livestock manure is the main source of infection of schi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12A61K39/00A61K48/00A61P33/12
CPCY02A50/30
Inventor 刘金明宋震宇孙毅石耀军李浩林矫矫
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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