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SiRNA (small interfering RNA) sequences of targeted inhibition capripoxvirus ORF095 genes

A technology of ORF095, sheep pox virus, applied in DNA/RNA fragmentation, gene therapy, recombinant DNA technology, etc., can solve the problem of no RNA interference method, etc.

Inactive Publication Date: 2013-01-02
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the research of targeting sheeppox virus based on RNA interference technology in the world at present, and there is no report on the research of RNA interference method on ORF095 protein.

Method used

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  • SiRNA (small interfering RNA) sequences of targeted inhibition capripoxvirus ORF095 genes
  • SiRNA (small interfering RNA) sequences of targeted inhibition capripoxvirus ORF095 genes
  • SiRNA (small interfering RNA) sequences of targeted inhibition capripoxvirus ORF095 genes

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Embodiment Construction

[0041] The present invention is explained in detail below in conjunction with embodiment.

[0042] 1. Preparation of Sequences

[0043] The four sequences designed by the present invention are respectively: the gene sequence of siRNA-61 is SEQ.NO.1; the gene sequence of siRNA-70 is SEQ.NO.2; the gene sequence of siRNA-165 is SEQ.NO.3; The gene sequence of -296 is SEQ.NO.4.

[0044] Send the designed sequence to Shanghai Gemma Pharmaceutical Technology Co., Ltd. for 5' end modification synthesis.

[0045] 2. Construction of pEGFP-ORF095 expression vector

[0046] Use the TaKaRa company DNA extraction kit to extract DNA from cell strains, and use the extracted and purified viral DNA as a template for amplification. The PCR reaction system is: 10×PCR Buffer (containing Mg2+) 5 μL, dNTPs 4 μL, template 1 μL, primer F 1 μL each of P and P, 0.5 μL of rTaq enzyme, and make up to 50 μL with ultrapure water. The reaction conditions are: 95°C for 5min; 94°C for 30s, 55°C for 30s, 72...

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Abstract

The invention discloses siRNA (small interfering RNA) sequences of targeted inhibition capripoxvirus ORF095 genes and application of the sequences. The sequences of the four targeted inhibition capripoxvirus ORF095 genes are respectively siRNA-61 with the gene sequence of SEQ.NO.1, siRNA-70 with the gene sequence of SEQ.NO.2, siRNA-165 with the gene sequence of SEQ.NO.3, and siRNA-296 with the gene sequence of SEQ.NO.4. The siRNA sequences of the four targeted inhibition capripoxvirus ORF095 genes can be applied to preparation of vaccine for inhibiting capripoxvirus and preparation of drugs for inhibiting capripoxvirus, or cultivation of sheep with anti-capripoxvirus virus.

Description

technical field [0001] The present invention relates to four siRNA sequences targeting to inhibit the ORF095 gene of sheeppox virus and the application of these sequences. Background technique [0002] Capripox (CP) includes sheep pox, goat pox and bovine skin pimple disease, wherein the former two are caused by sheep pox virus (Sheeppox virus, SPPV) and goat pox virus (Goatpox virus, GTPV) respectively. contact infectious diseases. The body temperature of sick animals rises, papules or nodules, blisters, visceral lesions and even death appear on the whole body, which has caused serious economic losses to the sheep industry in the world. The World Organization for Animal Health (OIE) stipulates it as an animal disease that must be reported by law, and my country lists it as a first-class animal infectious disease. The prevention and control of sheeppox has always been a research hotspot. [0003] ORF095 is a gene encoding virion packaging core protein, which is homologous...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113A61K48/00A61K31/20C12N15/85A01K67/027
Inventor 张强赵志荀吴国华颜新敏李健朱海霞崔立凡高顺平才学鹏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI