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Chimeric type porcine circovirus live vaccine C1-233 strain and constructing method thereof

A technology of porcine circovirus and its construction method, which is applied in the field of construction of recombinant porcine circovirus type 1 live vector vaccine strain, and can solve the problems of difficult accurate control of incomplete enzyme digestion conditions and low construction success rate

Active Publication Date: 2017-06-13
YANGZHOU UNIV
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  • Application Information

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Problems solved by technology

It is worth emphasizing that when the above-mentioned authors constructed PCV1-2, they used Kpn Ⅰ The incomplete digestion method realizes the construction of double copies of PCV1-2. After many years of verification in our laboratory, the incomplete digestion conditions in this method are difficult to accurately control, resulting in an extremely low construction success rate

Method used

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  • Chimeric type porcine circovirus live vaccine C1-233 strain and constructing method thereof
  • Chimeric type porcine circovirus live vaccine C1-233 strain and constructing method thereof
  • Chimeric type porcine circovirus live vaccine C1-233 strain and constructing method thereof

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Embodiment Construction

[0043] 1. Construction of recombinant porcine circovirus type 1 live vector vaccine strain (C1-233 strain) expressing porcine circovirus type 2b ORF2 gene:

[0044] The primer sequences designed for amplification are as follows:

[0045] Primer name Primer sequence enzyme Kpn I-F:

5'-TTT GGTACC CGAAGGCCGATT-3'

Kpn I

Kpn I-R:

5'-ATT GGTACC TCCGTGGATTGTTCT -3'

Kpn I

Nar I-F:

5'-GGT GGCGCC TCCTTGGATACGTCATCCTATAAAAGTG-3'

Nar I

Hpa I-R:

5'-GAA GTTAAC CCTAAATGAATAAAAATAAAAACCATTACG-3'

Hpa I

Psi I-F:

5'-AGG TTATAA GTGGTGGGGGGTCTTTAAGATTAA-3'

Psi I

Acl I-R:

5'-GGA AACGTT ACCGCAGAAGAAGACACC-3'

Acl I

[0046] Note: The underlined part is the recognition sequence of each endonuclease

[0047] 1. Construct pSK(+)-PCV1ΔORF2 with deletion of PCV1 ORF2 genome:

[0048] like Figure 1a As indicated, using the whole genome of PCV1 as a tem...

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Abstract

The invention provides a chimeric type porcine circovirus live vaccine C1-233 strain and a constructing method thereof, and belongs to the technical field of vaccine study production. The constructing method comprises the steps of screening of PCV1 and PCV2 strains, construction of a recombined porcine circovirus 1 type live vector vaccine strain which expresses a porcine circovirus 2b type ORF2 gene, recombination of viral nucleic acid, and composition characteristics and biological characteristics of an amino acid sequence. Finally a recombined porcine circovirus 1 type live vector vaccine strain (C1-233 strain) which can be used for industrialized production and expressing the porcine circovirus 2b type ORF2 gene is obtained.

Description

technical field [0001] The invention belongs to the technical field of vaccine research and production, in particular to the construction technology of a recombinant porcine circovirus type 1 live vector vaccine strain expressing porcine circovirus type 2b ORF2 gene. Background technique [0002] Porcine circovirus type 2 (PCV2) seriously endangers the healthy development of the pig industry in the world, and PCV2 is widely distributed all over the world. Porcine circovirus type 1 (PCV1) was first discovered in 1974 by the German scholar Tischer from the PK15 passage cell line as a cell pollutant [Tischer I, Rasch R, Tochtermann G, et al .Characterization of papovavirus and picornavirus-like particles in permanentpig kidney cell lines[J].Zentralbl Bakteriol Orig A, 1974, 226 (2): 153-167.], although prevalent in all parts of the world, so far have not been found to have Disease-causing ability. PCV2 was subsequently isolated from sick pigs. It is very similar to PCV1, but ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/85C12N15/66C12R1/93
CPCC12N7/00C12N15/66C12N15/85C12N2750/10021C12N2750/10051C12N2800/107
Inventor 高崧刘秀梵高清清王小波高如一何洪波李基棕樊铭玉程清如
Owner YANGZHOU UNIV
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