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Porcine teschovirus DBN-10021 strain complete genome sequence, primer for determining same, and application thereof

A technology of DBN-10021 and porcine Jieshen virus, which is applied in the field of molecular biology, can solve the problems affecting the determination of the full-length sequence of the porcine Jieshen virus genome, the large variation of the porcine Jieshen virus genome, and the difficulty in designing primers

Active Publication Date: 2012-11-14
兆丰华生物科技(南京)有限公司 +1
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Problems solved by technology

Many studies in my country have shown that the positive rate of porcine Jieshen virus in pig herds is relatively high, but in my country there are only research reports on porcine Jieshen virus type 2 and porcine Jieshen virus type 8 genomes, the main reason is that the porcine Jieshen virus genome The variation is large, and it is difficult to design primers, which directly affects the determination of the full-length sequence of the porcine Jieshen virus genome

Method used

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  • Porcine teschovirus DBN-10021 strain complete genome sequence, primer for determining same, and application thereof
  • Porcine teschovirus DBN-10021 strain complete genome sequence, primer for determining same, and application thereof
  • Porcine teschovirus DBN-10021 strain complete genome sequence, primer for determining same, and application thereof

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Embodiment 1

[0020] Cloning of the full genome sequence of porcine Jieshen virus DBN-10021 strain of embodiment 1

[0021] (1) Extraction of viral RNA

[0022] The porcine Jieshen virus DBN-10021 strain (CGMCC No.5573) was taken and passaged by PK15 cells, the venom of the fifth generation cells was collected, frozen and thawed three times, and then centrifuged at 12000r / min at 4°C for 5 minutes. Take 250 μl of the virus suspension and put it in a 1.5ml EP tube, add 0.75ml Transzol (Beijing Quanshijin Biotechnology Co., Ltd.), mix well; then add 200 μl of chloroform, shake vigorously for 15 seconds, and incubate at room temperature for 3 minutes. Centrifuge at 12000r / min at 4°C for 10 minutes. After centrifugation, transfer the upper aqueous phase to a new 1.5ml EP tube, add 0.5ml isopropanol, invert to mix, and incubate at room temperature for 10 minutes. Centrifuge at 12000r / min at 4°C for 10 minutes, discard the supernatant and keep the precipitate. Add 1ml of 75% ethanol (prepared w...

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Abstract

The invention discloses a porcine teschovirus DBN-10021 strain complete genome sequence, a primer for determining same, and application thereof. The porcine teschovirus DBN-10021 strain complete genome sequence provided by the invention is shown as SEQ ID No. 1. The primer used for amplifying the porcine teschovirus DBN-10021 strain complete genome sequence comprises a primer of which the sequence is shown as SEQ ID No. 2 to 13. The virus genome sequence is divided into six fragments, and the corresponding cDNA (complementary deoxyribonucleic acid) is amplified by an RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) method; target fragments can be effectively amplified, and the complete genome sequence is provided; and the invention is favorable for studying classification, evolution, pathogenicity and molecular epidemiology of virus and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a method for determining the whole genome sequence of Jieshen virus type 4, in particular to a method for measuring the whole genome sequence of porcine Jieshen virus DBN-10021 strain. Background technique [0002] Porcine teschovirus (Porcine teschovirus, porcine teschovirus) can cause porcine poliomyelitis, reproductive disorders, pneumonia, diarrhea, pericarditis and myocarditis, skin damage and asymptomatic and other clinical manifestations. There are many genotypes of Jieshen virus, at least 11 genotypes currently exist, and there are also large differences in the genomes of different isolates of the same genotype. Many studies in my country have shown that the positive rate of porcine Jieshen virus in pig herds is relatively high, but in my country there are only research reports on porcine Jieshen virus type 2 and porcine Jieshen virus type 8 genomes, the main reason is that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C12N15/11C12Q1/70C12Q1/68
Inventor 宫晓文王美君张志榜焦连国王贵华赵亚荣
Owner 兆丰华生物科技(南京)有限公司
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