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A kind of cpg-odn with specific immunostimulatory effect on prrsv and application thereof

An immunostimulatory and specific technology, applied in the field of CpG-ODN, to achieve the effect of reducing production cost, reducing morbidity, improving survival rate and breeding efficiency

Active Publication Date: 2021-06-15
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the immunostimulatory activity of two immune activation units (-CG-motif and -GTCGTT-motif) at the same time.

Method used

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  • A kind of cpg-odn with specific immunostimulatory effect on prrsv and application thereof
  • A kind of cpg-odn with specific immunostimulatory effect on prrsv and application thereof
  • A kind of cpg-odn with specific immunostimulatory effect on prrsv and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0028] 1. Screening of specific CpG-ODN fragments

[0029] Screening method: According to conventional techniques, CpG-ODN fragments were directional screened, and the results are shown in Table 1 below.

[0030] Table 1 Screened CpG-ODN fragments

[0031]

[0032] The CpG-ODN fragments screened above are screened based on the research of related sequences by relevant personnel, for example:

[0033] References for C274 are "Marshall, J.D., Higgins, D., Abbate, C., et al. (2004). Polymyxin B enhances ISS-mediated immune responses across multiple species. Cellular Immunology, 229, 93–105." and "Teleshova , N., Kenney, J., Williams, V., et al. (2006). CpG-C ISS-ODN activation of blood-derived B cells from healthy and chronic immunodeficiency virus-infected macaques. Journal of Leukocyte Biology, 79, 257–267 "."

[0034]BCG-A4a reference is "Yamamoto, S., Yamamoto, T., Nojima, Y., et al. (2002). Discovery of immunostimulatory CpG-DNA and its application to tuberculosis vacc...

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Abstract

The present invention relates to a CpG-ODN with specific immunostimulatory effect on PRRSV and its application. By screening out 11 CpG-ODN fragments, the screened 11 CpG-ODN fragments are connected end-to-end to form a composition containing a number of CpG-ODN fragments. A ‑dinucleotide is a DNA sequence of a CpG motif, named "Multi‑CpG". A DNA sequence of the same length in the above "Multi-CpG" will be replaced with a DNA sequence of -GTCGTT- as a CpG motif, and named "Multi-CpG-Mix". Two "Multi-CpG" and one "Multi-CpG-Mix" were connected end-to-end to obtain a DNA sequence containing two immune activation units, which was named "Multi-CpG-Mixture". The "Multi-CpG-Mixture" was inserted into the multi-copy cloning vector Puc19-derived vector FVX through the two restriction sites of XbaI and EcoRI, and the recombinant plasmid FVX-Multi-CpG-Mixture was constructed by means of gene synthesis. It can not only induce the activation of natural immune function in pigs, but also activate the acquired immune function of PRRSV in pigs, improve the induction of immune response to PRRSV in pigs, reduce the incidence of PRRSV, and reduce the cost of pig breeding.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a CpG-ODN with specific immunostimulatory effect on PRRSV and its application. Background technique [0002] Porcine reproductive and respiratory syndrome virus (PRRSV) is the pathogenic factor of porcine reproductive and respiratory syndrome (PRRS), and it is a new RNA virus infectious disease virus that causes reproductive disorders in sows, respiratory symptoms and high mortality in newborn piglets. PRRSV was first discovered in the United States in 1987. In 1991, the Dutchman Wensvoot isolated the virus from sick piglets and sows for the first time, and then Germany, the United States, and the United Kingdom also isolated the virus. At present, the virus has spread throughout North America and Europe, and has spread globally, and epidemic infections have occurred successively in various places, causing serious economic losses to the pig industry. Although porcine repr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/117C12N15/85A61K48/00A61P31/14
CPCA61K48/005A61P31/14C12N15/117C12N15/85C12N2310/17C12N2320/30C12N2800/107
Inventor 冯璥李涛何文彬吕雪梅
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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