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Sequence siRNA-70 (small interfering Ribonucleic Acid) for targeted inhibition of capripoxvirus ORF095 gene

A technology of ORF095 and sheep pox virus, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problem of no RNA interference method

Inactive Publication Date: 2012-11-28
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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  • Sequence siRNA-70 (small interfering Ribonucleic Acid) for targeted inhibition of capripoxvirus ORF095 gene
  • Sequence siRNA-70 (small interfering Ribonucleic Acid) for targeted inhibition of capripoxvirus ORF095 gene
  • Sequence siRNA-70 (small interfering Ribonucleic Acid) for targeted inhibition of capripoxvirus ORF095 gene

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Embodiment Construction

[0042] The present invention is explained in detail below in conjunction with embodiment.

[0043] 1. Preparation of Sequences

[0044] 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.

[0045] Send the designed sequence to Shanghai Gemma Pharmaceutical Technology Co., Ltd. for 5' end modification synthesis.

[0046] 2. Construction of pEGFP-ORF095 expression vector

[0047] DNA was extracted from cell strains using a DNA extraction kit from TaKaRa Company, and the purified viral DNA was used as a template for amplification. The PCR reaction system was: 10×PCR Buffer (containing Mg2+) 5 μL, dNTPs 4 μL, template 1 μL, primers 1 μL each of F and P, 0.5 μL of rTaq enzyme, and make up to 50 μL with ultrapure water. The reaction conditions were: 95°C for 5 min; 30 cycles of 94°C for 30 s, ...

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Abstract

The invention discloses a sequence siRNA-70 (small interfering Ribonucleic Acid) for targeted inhibition of a capripoxvirus ORF095 gene and application of the sequence.

Description

Technical field [0001] The invention involves four targeted SiRNA sequences that inhibit the ORF095 gene, as well as the purpose of these sequences. Background technique [0002] Capripox (CP) includes the skin's skin disease of cotton, acne, and beef. Among them, the first two are sheep acne virus (SPPV) and Goatpox virus (GTPV), respectively.Contact infectious diseases.The physical temperature of the diseased animal is rising, and pimples or nodules, blisters, internal organs, or even death appear in the whole body, causing serious economic losses to the sheep industry in the world.The World Animal Health Organization (OIE) stipulates it as an animal epidemic that must be reported on the statutory, and my country lists it as a type of animal infectious disease.The prevention and control of sheep acne have always been a hot spot for research. [0003] ORF095 is the core protein gene encoding the core protein gene of virus particle packaging (Accession No.170726) of MonkeyPox Vir...

Claims

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Application Information

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IPC IPC(8): C12N15/113
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