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Recombinant adenovirus of porcine reproductive and respirator syndrome virus and porcine Circovirus, and vaccine

A technique for respiratory syndrome and recombinant adenovirus, applied in virus/bacteriophage, biochemical equipment and methods, applications, etc.

Inactive Publication Date: 2006-07-12
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

The recombinant adenovirus has been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee

Method used

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  • Recombinant adenovirus of porcine reproductive and respirator syndrome virus and porcine Circovirus, and vaccine
  • Recombinant adenovirus of porcine reproductive and respirator syndrome virus and porcine Circovirus, and vaccine
  • Recombinant adenovirus of porcine reproductive and respirator syndrome virus and porcine Circovirus, and vaccine

Examples

Experimental program
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Embodiment Construction

[0026] (1) Construction of genetically engineered body-PRRSV and PCV recombinant adenovirus (rAd-Cap-GP5)

[0027] 1 Construction and identification of recombinant plasmid pShuttle-Cap-GP5

[0028] 1.1 Amplification of Cap protein gene:

[0029] 1.1.1 Primers: Using the PCV2-TJ (serial number AY181946) strain gene sequence in GenBank as a template, a pair of primers were designed with the software PrimerPremier 5.0, and a Kpn I restriction site was added to the 5' ends of both primers. Primer 1: 5-TTCggT ACC AgC TAT gAC gTA TCC AAg-3, Primer 2: 5-gCA ggT ACC gTT AAg Tgg ggg gTC TTT-3. The size of the gene fragment amplified by the primers is 718bp.

[0030]1.1.2 DNA extraction: Freeze and thaw the PCV2-PK15 cell culture three times, transfer to a 1.5ml centrifuge tube, centrifuge at 12,000 rpm for 5 minutes, and take the supernatant; add an equal volume of chloroform, mix well, centrifuge at 12,000 rpm for 10 minutes, and take The supernatant was repeatedly extracted three ...

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Abstract

The invention relates to a pig breeding and breathing complex virus (PPRSV) and pig 2-type loop virus (PCV 2) series gene recombination adenovirus and vaccine in the field of high and new biotechnology. It uses PCR technology to clone the Cap protein gene into the plasmid carrier pShuttle-CMV-GP5 by open type reading rule and transforms tobacillus coli BJ5183strain with cage carrier pAdEasy-1 to capture the recombination plasmid; it uses recombination plasmid HEK293-A cell to capture recombination adenovirus and purifies it to express the recombination adenovirus rAd-Cap-GP5 of PRRSV GP5protein and PCV-2 Cap protein.

Description

1. Technical field [0001] The porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus tandem gene recombinant adenovirus and vaccine of the present invention belong to the high-tech field of biotechnology, and are specially used to strengthen the protection of pigs against porcine reproductive and respiratory syndrome and type 2 porcine circovirus Immunoprotective effect and reduce the economic loss of pig farms. 2. Technical background [0002] Recombinant adenovirus genetic engineering technology has been widely used in human gene therapy, and many recombinant adenoviruses used in human gene therapy have entered the third phase of clinical trials. Because the recombinant adenovirus has a wide range of hosts, can infect many types of cells, can express human and non-human proteins, and will not insert into the host chromosome, has a low reverse mutation rate, and can be replicated to a high titer. The GP5 protein is a glycoprotein of about 25 kD ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/861C12N15/33A61K39/235
Inventor 姜平王先炜李玉峰
Owner NANJING AGRICULTURAL UNIVERSITY
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