Recombinant virus expressing PCV2 codon optimized ORF1 and ORF2 tandem gene

A technology of codon optimization and recombinant virus, which is applied in the fields of genetic engineering vaccines and bioengineering, to achieve the effects of increased expression, good immunogenicity and protection, and high safety

Inactive Publication Date: 2013-09-04
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that there is genetic diversity in PCV2 in my country, so vaccines designed b

Method used

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  • Recombinant virus expressing PCV2 codon optimized ORF1 and ORF2 tandem gene
  • Recombinant virus expressing PCV2 codon optimized ORF1 and ORF2 tandem gene
  • Recombinant virus expressing PCV2 codon optimized ORF1 and ORF2 tandem gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0046] 1. Construction of recombinant baculoviruses expressing foreign genes

[0047] 1) Optimization and synthesis of PCV2 ORF2 gene

[0048] By comparing and analyzing all the sequences of PCV2 in GenBank, the consensus sequences of PCV2 ORF1 and ORF2 expressed amino acids were obtained, and the PCV2 ORF1 and ORF2 genomes were transformed into partial codons in insect cells through codon optimization, and according to specific conditions (GC content, Avoid repeated sequences, etc.) to make appropriate adjustments, if the codon with the highest frequency cannot be used, select the codon with the second highest frequency. And optimize and transform the rare codons relative to baculovirus. Artificially synthesized codon-optimized PCV2 ORF1 and ORF2 tandem genes, the gene is named YHsfORF1-2, and its sequence is (SEQ.ID.NO.1):

[0049] 1 ATGCCCTCAAAGAAGAACGGTAGAAGCGGACCACAGCCACACAAGAGATG

[0050] 51 GGTTTTTCACACTCAAACAACCCCTCAGAAGACGAGCGTAAGAAGATCAGAG

[0051] 101 AGTTGCCAAT...

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Abstract

The invention relates to a recombinant virus of a PCV2 codon optimized ORF1 and ORF2 tandem gene, and belongs to the biopharmaceutical field. The artificially-synthesized codon optimized PCV2 ORF1 and ORF2 gene is named YHsfORF1-2. The YHsfORF1-2 is cloned to a baculovirus expression vector pFastBacHTA to obtain a recombinant plasmid pFBHYHsfORF1-2; and the recombinant plasmid is transfected to an Sf9 cell to obtain a recombinant baculovirus vFBHYHsfORF1-2. In the recombinant baculovirus vFBHYHsfORF1-2, ORF1 and ORF2 proteins can be successfully efficiently expressed, and can form virus-like particles. The invention also relates to an application of the recombinant virus in vaccines. Animal tests prove that inactivated vaccines prepared through using the recombinant virus can stimulate the body to generate efficient immune response and have a good immune protection effect on PCV2.

Description

1. Technical field [0001] The invention relates to a recombinant virus expressing PCV2 codon-optimized ORF1 and ORF2 tandem genes, which belongs to the field of bioengineering, in particular, relates to the optimization of porcine circovirus ORF1 and ORF2 codons, the acquisition of optimized sequence recombinant baculovirus and The genetic engineering vaccine prepared by it belongs to the field of genetic engineering vaccines. 2. Background technology [0002] Porcine circovirus disease (PCVD) is an infectious disease of porcine caused by porcine circovirus type 2 (PCV2). loss. Many scholars believe that porcine circovirus disease can cause multisystemic wasting syndrome (PMWS) and porcine dermatitis nephrotic syndrome (PDNS) after weaning, and can also cause reproductive disorders in sows, congenital tremor (CT) in newborn piglets, etc. It is also one of the primary pathogens of porcine respiratory disease syndrome (PRDC). At present, PCV2 infection is common in pig farm...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/34C12N15/866A61K39/12A61P31/20C12R1/93
Inventor 王小敏何孔旺汪伟周忠涛俞正玉倪艳秀温立斌张雪寒郭容利吕立新李彬周俊明茅爱华叶青周萍沈江萍
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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