Peste des petits ruminants virus CH/GDDG/2014 strain whole genome sequence and amplification primer pair thereof

A PPR and whole genome technology, applied in the field of molecular biology, can solve the problems of no public display, affecting the pathogenicity of the virus growth curve, unfavorable for the new type of PPR

Active Publication Date: 2015-12-02
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

At present, although the whole genome sequence of the new type 4 PPR virus has been reported, there is no public indication of how it was amplified, which is not conducive to the construction of a reverse genetic system for the new type of PPR virus in my country. At the same time, it will further affect our understanding Biological characteristics such as growth curve, virus titer and pathogenicity of the virus

Method used

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  • Peste des petits ruminants virus CH/GDDG/2014 strain whole genome sequence and amplification primer pair thereof
  • Peste des petits ruminants virus CH/GDDG/2014 strain whole genome sequence and amplification primer pair thereof
  • Peste des petits ruminants virus CH/GDDG/2014 strain whole genome sequence and amplification primer pair thereof

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Embodiment Construction

[0018] The present invention will be further described below in conjunction with specific embodiments, and its operation steps will be further clarified.

[0019] Optimized embodiment:

[0020] (1) Extraction of viral RNA

[0021] Take goat lung samples identified as positive for Peste des petits ruminants virus by RT-PCR, grind, homogenate, dilute at 1:2, freeze and thaw repeatedly 3 times, and extract RNA according to the virus RNA extraction kit (Axygen Company) , and finally, use 50-100 μL RNase-free double-distilled water for RNA elution, and store in a -20°C refrigerator;

[0022] (2) Primer design and synthesis

[0023] Through the comparison of the entire genome sequences of multiple genotypes of Peste des petits ruminants virus, the PrimerPremier5.0 software was used to select conserved sequences as primers. There are overlapping sequences of different lengths between adjacent amplified fragments, which is convenient for later sequence splicing . A total of 11 pai...

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Abstract

The invention discloses a peste des petits ruminants virus CH / GDDG / 2014 strain whole genome sequence and an amplification primer pair thereof. The virus genome sequence is divided into 11 segments, corresponding cDNA is amplified by using a one-step RT-PCR method, and amplified PCR segments are cloned onto a pGM-T carrier for nucleic acid sequencing. By the virus genome sequence, target segments can be amplified effectively; the whole genome sequence is provided, and studying of genetics, molecular epidemiology and reverse genetics of peste des petits ruminants virus is facilitated.

Description

technical field [0001] The patent of the present invention belongs to the field of molecular biology, and relates to a method for determining the whole genome sequence of Peste des petits ruminants genotype 4, in particular to a method for determining the whole genome sequence of Peste des petits ruminants virus CH / GDDG / 2014 strain. Background technique [0002] Peste des petits ruminants is a severe infectious disease that seriously endangers the health of goats, sheep, wild ruminants and other animals. It can cause various clinical symptoms such as pneumonia, diarrhea, stomatitis and abortion in infected animals. The World Organization for Animal Health lists it as category A animal disease. There are 4 genotypes of Peste des petits ruminants, and there are large differences between the genomes of different genotypes or strains of the same genotype. In 2007 and 2013, two waves of Peste des Petits Ruminants broke out in my country’s Tibet and Xinjiang, especially the secon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12N15/40
CPCC12Q1/686C12Q1/70C12Q1/701
Inventor 翟少伦魏文康吕殿红温肖会
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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