Biological preparation for resisting against replication and infection of foot-and-mouth disease virus and preparation method as well as application thereof

An anti-foot-and-mouth disease virus and biological agent technology, which is applied in the directions of biochemical equipment and methods, antiviral agents, and botanical equipment and methods, can solve the problems of difficulty in achieving the protective effect of mutant strains, and achieves inhibition of foot-and-mouth disease virus replication and inhibition. The effect of anti-foot-and-mouth disease infection

Active Publication Date: 2012-06-27
SOUTH CHINA AGRI UNIV
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  • Abstract
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Problems solved by technology

However, it takes at least seven days for the vaccine to produce a protective effect. At the same time, due to the large number of FMDV serotypes, there is almost no cross-protection betwee

Method used

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  • Biological preparation for resisting against replication and infection of foot-and-mouth disease virus and preparation method as well as application thereof
  • Biological preparation for resisting against replication and infection of foot-and-mouth disease virus and preparation method as well as application thereof
  • Biological preparation for resisting against replication and infection of foot-and-mouth disease virus and preparation method as well as application thereof

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Embodiment 1

[0060] The present invention is based on O-type FMDV isolated from different ages and regions, selects highly conserved sequences of FMDV 2B and VP1 genes, designs and synthesizes siRNA, clones them into the PSIREN-shuttle shuttle vector, and constructs recombinant plasmids pShuttle-2B and pShuttle-VP1. The specific process as follows:

[0061] (1) Dilute the transcription template of siRNA-VP1 and the transcription template of siRNA-2B to the same concentration (5pmol / μL) with deionized water, and mix the two complementary oligonucleotides (5μL each) Then put it on the PCR amplification instrument, treat at 95°C for 2min, and then cool down to normal temperature naturally for processing. The double-stranded siRNA oligonucleotide fragments formed after treatment were respectively ligated with pSIREN-Shuttle (Clontech Company) after double digestion with BamHI and EcoRI (the ligation reaction was operated according to the instructions of T4 DNA ligase, and the screening of posi...

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Abstract

The invention discloses a biological preparation for resisting against replication and infection of a foot-and-mouth disease virus and a preparation method as well as application thereof. Active parts of the biological preparation are siRNA-VP1 and siRNA-2B; a nucleotide sequence of a sense strand of a transcription template of the siRNA-VP1 is shown as SEQ ID NO. 1; and a nucleotide sequence of a sense strand of a transcription template of the siRNA-2B is shown as SEQ ID NO. 3. In the invention, by connecting the transcription template of the siRNA-VP1 and the transcription template of the siRNA-2B with U6 promoter, human replicated defect 5 type adenovirus plasmid carrier pAdeno-X is cloned to obtain plasmids which can be transcribed and connected in series respectively; and next, cell packaging is performed through packaging to obtain recombinant adenovirus Adeno-2B-VP1. Experiments prove that the prevention and control effect of the recombinant adenovirus Adeno-2B-VP1 is superior to the effect obtained by independently applying the recombinant adenovirus siRNA-2B and the recombinant adenovirus Adeno-2B-VP1 or applying the recombinant adenovirus siRNA-2B and the recombinant adenovirus Adeno-2B-VP1 in a combined way. The biological preparation can be used for suppressing replication of the foot-and-mouth disease virus and resisting against the infection of the foot-and-mouth disease.

Description

technical field [0001] The invention belongs to the technical field of foot-and-mouth disease prevention and control, and particularly relates to a biological preparation for resisting replication and infection of foot-and-mouth disease virus, a preparation method and application thereof. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease of artiodactyls caused by foot-and-mouth disease virus (FMDV). The infection rate of the disease is high, and the highest incidence rate after infection can reach 100%. There are more than 70 susceptible animals, among which important economic livestock species such as pigs, cattle, and sheep are all susceptible. In view of the fact that FMD not only reduces animal production performance and causes huge direct economic losses, but also affects the international trade of animals and animal products, so the indirect losses caused are even greater, and it will also affect a cou...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/713A61K39/235A61K35/76C12N15/113C12N15/861C12N7/01A61P31/14A61K35/761
Inventor 陈金顶李银光赵明秋徐艳芳刘文俊代曼曼
Owner SOUTH CHINA AGRI UNIV
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