Brucella deoxyribonucleic acid (DNA) vaccine as well as construction method and application thereof

A technology of DNA vaccine and Brucella, which is applied in the field of vaccine application, Brucella DNA vaccine and its construction, can solve the problems of protection effect, safety and production cost that are difficult to overcome, and achieve long duration of immunity, Good safety, simple preparation and effective effect

Inactive Publication Date: 2012-11-14
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there are many types of Brucella vaccines and they have played an important role in the immune prevention and control of brucellosis in the world, so far no traditional Brucella vaccine has been widely or persistently used in the world. Various types of vaccines have different degrees of contradictory and insurmountable shortcomings in terms of protective effect, safety and production cost.

Method used

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  • Brucella deoxyribonucleic acid (DNA) vaccine as well as construction method and application thereof
  • Brucella deoxyribonucleic acid (DNA) vaccine as well as construction method and application thereof
  • Brucella deoxyribonucleic acid (DNA) vaccine as well as construction method and application thereof

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Embodiment Construction

[0031] The present invention is further described below in conjunction with embodiment. The examples are intended to illustrate the present invention, but not to limit the present invention in any form.

[0032] The present invention clones the P39 and 18KD genes from the whole genome of Brucella abortus type 2 CVCC 12 strains, obtains the P39-IRES-18KD fragment through the pQCXIX reverse transcription vector, and then directional clones it into the eukaryotic expression vector pcDNA3. A DNA vaccine was constructed in 1(+).

[0033] Its specific construction method is:

[0034] 1. Cloning of P39 and 18KD genes

[0035] Get 12 strains of Brucella abortus type 2 CVCC (can be purchased from China Veterinary Drug Administration), use Wizard Genomic DNA Purification Kit (Promega company) is from the bacterial total DNA that Brucella abortus type 2 CVCC 12 strains extracts, with this as template, with the sequence that table 1 provides as primer, by polymerase chain reaction, am...

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Abstract

The invention discloses a Brucella deoxyribonucleic acid (DNA) vaccine and a construction method thereof as well as the application of the vaccine. The Brucella DNA vaccine disclosed by the invention contains Brucella P39 and 18KD genes which can also be co-expressed in eukaryotic cells by internal ribosome entry site (IRES) mediation. The Brucella DNA vaccine disclosed by the invention has an SEQ1 gene sequence. According to the Brucella DNA vaccine and the construction method thereof, which are disclosed by the invention, the total DNA of Brucella abortus 2 type CVCC 12 is taken as a template, a polymerase chain reaction is carried out by using P39S primers SEQ2 and SEQ3 and 18KDS primers SEQ4 and SEQ5 respectively so as to obtain a target gene, and then the target gene is converted into a JM109 competent cell after being cleaved so as to obtain positive plasmid named as pQC-P39 / 18KD. Then, the pQC-P39 / 18KD and pcDNA3.1 plasmid are connected by using a T4DNA ligase after being cleaved, and afterwards are converted into E.coli DH5 alpha so as to obtain pcDNA-P39 / 18KD recombinant plasmid.

Description

technical field [0001] The present invention relates to a DNA vaccine and its construction method, as well as the application of the vaccine. Specifically, the present invention relates to a Brucella DNA vaccine, its construction method, and the application of the vaccine. Background technique [0002] Brucellosis is a zoonotic infectious disease caused by Brucella spp., also known as Malta fever or undulate fever, referred to as brucellosis. The World Organization for Animal Health (OIE) lists it as a B-type animal disease, and my country also lists it as a B-type animal disease, which directly affects the development of animal husbandry, seriously threatens human health, and causes huge economic losses (Cai Baoxiang, 2001). Brucellosis is caused by its unique pathogenic mechanism and the characteristics that it is difficult to cure after intracellular parasitic infection. In addition to large-scale culling, the application of vaccine prevention has always been the most eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/10A61K48/00A61P31/04C12N15/85C12R1/01
Inventor 蔺国珍邱昌庆郑福英景志忠
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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