Construction, rescue and application of infectious clone of porcine reproductive and respiratory syndrome virus type 1 isolate

A technology for respiratory syndrome and infectious cloning, which is applied in the field of bioengineering, can solve problems that have not been reported, achieve the effect of simple and convenient method, and overcome easy degradation

Inactive Publication Date: 2020-09-18
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the PRRSV reverse genetic manipulation platform has been established in previous studies, there is no report on the reverse genetic manipulation platform based on the PRRSV1 wild strain in my country.

Method used

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  • Construction, rescue and application of infectious clone of porcine reproductive and respiratory syndrome virus type 1 isolate
  • Construction, rescue and application of infectious clone of porcine reproductive and respiratory syndrome virus type 1 isolate
  • Construction, rescue and application of infectious clone of porcine reproductive and respiratory syndrome virus type 1 isolate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1—Porcine reproductive and respiratory syndrome virus type 1 attenuated strain HLJB1 genome full-length cDNA clone construction and rescue

[0028] 1.1 Construction of pACYC177-New vector

[0029] Referring to the previously reported infectious clone construction method and improving on the original basis, a method for constructing the HLJB1 infectious clone platform was designed. DNAMAN software was used to compare and analyze the whole genome sequence of HLJB1, and five single restriction sites of SgsI, Pfl23II, BglII, Bsp1407I and XbaI were selected for segmental insertion of the full length of the gene when the HLJB1 infectious cloning platform was constructed. Such as figure 1As shown in the schematic diagram of vector transformation, the low-copy plasmid pACYC177 was selected as the original vector, and a vector containing the CMV promoter, four corresponding restriction sites (SgsI, Pfl23II, BglII, Bsp1407I and XbaI) and bovine growth hormone (BGH) polya...

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Abstract

The invention relates to construction, rescue and application of infectious clone of porcine reproductive and respiratory syndrome virus type 1 isolate, and provides an infectious clone virus of porcine reproductive and respiratory syndrome virus type 1 (PRRSV1) wild strain. A key genetic operation platform is provided for researching Marc-145 cell adaptability determinants of the PRRSV1. Meanwhile, aiming at the practical problem that the PRRSV1 vaccine is lacked in China at present, the PRRSV1 wild strain infectious cloning platform disclosed by the invention can also be used for developinga safer and more efficient novel PRRSV1 gene engineering vaccine.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the construction, rescue, transformation and application of an infectious clone of a porcine reproductive and respiratory syndrome virus type 1 strain. Background technique [0002] Porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV) is widespread in pig raising countries all over the world. PRRSV infection mainly caused abortion in sows and respiratory symptoms in the whole herd. PRRSV can be divided into two genotypes (PRRSV1 and PRRSV2) and multiple genotypes. In 1995, PRRSV2 appeared for the first time in my country and spread rapidly throughout the country. In 2006, my country had an outbreak of "swine hyperthermia" caused by a highly pathogenic PRRSV2 variant (HP-PRRSV). In addition, in recent years, wild strains of PRRSV1 have also been isolated in Hong Kong, Inner Mongolia, Heilongjiang, Liaoning, Fuji...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/40C12N7/00A61K39/12A61P31/14C12R1/93
CPCA61K39/12A61K2039/5254A61K2039/552A61P31/14C12N7/00C12N15/85C12N2770/10021C12N2770/10034C12N2770/10052C12N2800/107
Inventor 陈南华李树滨朱建中
Owner YANGZHOU UNIV
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