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Foot-and-mouth disease virus resistant recombinant live bacteria and its preparation method

A technology of anti-foot-and-mouth disease virus and live bacteria vaccine, which is applied in the field of anti-foot-and-mouth disease virus recombinant live bacteria vaccine and its preparation, can solve problems such as the difficulty of foot-and-mouth disease prevention, and achieve the effect of convenience

Inactive Publication Date: 2006-02-08
FUDAN UNIV
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Problems solved by technology

Prevention of foot and mouth disease is quite difficult

Method used

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  • Foot-and-mouth disease virus resistant recombinant live bacteria and its preparation method
  • Foot-and-mouth disease virus resistant recombinant live bacteria and its preparation method
  • Foot-and-mouth disease virus resistant recombinant live bacteria and its preparation method

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Embodiment Construction

[0018] The DNA sequences at positions 62ヘ120 and 423ヘ480 in the VP1 gene of O-type FMDV were chemically synthesized in series to form a primary structure of 423ヘ480-62ヘ120-423ヘ480. This fragment was cloned into the eukaryotic vector pCDM8 to construct pBO1.

[0019] Plasmid pBO1 was then treated with CaCl 2 Transformed into intermediate host attenuated Salmonella typhimurium LB5010. The plasmid was extracted from LB5010, and transformed into attenuated Salmonella choleraesuis C500 by electric shock method. Pick the attenuated Salmonella choleraesuis C500 strain in 3mL LB liquid medium, shake overnight at 37°C 200r / min; take 250μL of the bacterial liquid, inoculate it in 25mL LB liquid medium, and grow to OD at 37°C 200r / min 600 =0.6-0.7 (about 2-2.5h); 4°C, 4000r / min centrifuge for 10min to collect the bacteria; 0°C, wash with 10% glycerol, 4°C, 4000r / min for 10min to collect the bacteria, 3 times; the last wash Then suspend in 1mL of 10% glycerol, pack in 200μL per tube, a...

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Abstract

Disclosed is a recombinant live vaccine against foot-and-mouth disease virus delivered by Attenuated Salmonella choleraesuis, whose preparation consists of synthesizing the DNA sequence encoding the foot-and-mouth disease virus antigen epitope genes through chemical synthesis method, linking into complete antigen genes on the DNA horizontally to form fusion genes, inserting the fusion genes into expression plasmid vectors to construct eukaryon expression plasmid pB01, passing the eukaryon expression plasmid pB01 through intermediate host of samonella typhimurium LB5010, electrically transforming into Attenuated Salmonella choleraesuis, thus constructing the recombinant live vaccine pB01 / S.cho against foot-and-mouth disease virus.

Description

technical field [0001] The invention belongs to the technical field of foot-and-mouth disease virus prevention and control, and in particular relates to an anti-foot-and-mouth disease virus recombinant live bacterin using attenuated Salmonella choleraesuis C500 (piglet paratyphoid live bacterin) as a carrier and a preparation method thereof. technical background [0002] Foot-and-mouth disease (FMD) is the most serious livestock infectious disease in the world today, mainly harming pigs, cattle, sheep and other cloven-hoofed animals. For many years, foot-and-mouth disease has broken out and become popular in a large scale all over the world, causing huge economic losses to animal husbandry. According to immunogenicity, FMD virus has 7 serotypes and more than 65 subtypes, including A, O, C, SAT I, SAT II, ​​SAT III and Asia I. Prevention of foot-and-mouth disease is quite difficult. At present, there are two main ways. The first way is to inactivate the virus through physic...

Claims

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Application Information

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IPC IPC(8): C12N1/21A61K39/112C12N15/63C12N15/09
Inventor 刘明秋严维耀郑兆鑫
Owner FUDAN UNIV
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