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Research of Actinobacillus Pleuropneumoniae Bacterial Ghost Loading of Pasteurella Multocida Outer Membrane Protein (Omp) H DNA Vaccine

Author: SunYing
Tutor: JinTianMing
School: Tianjin Agricultural
Course: Preventive Veterinary Medicine
Keywords: App Bacterial ghost Pasteurella multocida DNA vaccines OmpH antigen
CLC: S858.28
Type: Master's thesis
Year: 2011
Downloads: 71
Quote: 0
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Abstract


Porcine contagious pleuropneumonia farms common bacterial respiratory diseases , lung disease often has mixed or secondary infection , can do great damage to the pig industry . Production practices , vaccination is the best way to combat the disease . Commonly inactivated vaccine for heterologous serotype strains infected with a protective effect , unable to prevent bacterial colonization , to bring some difficulties for the production practice . Bacterial ghost vaccine is a novel form of a vaccine , the preparation process , the membrane surface antigen intact , which can effectively induce humoral immunity and cellular immunity . Therefore , bacteria shadow both as a vaccine carrier have extensive use value . This test the phage PhiX174 genome and the Staphylococcus aureus genome as a template by PCR amplification of the lysis protein E gene and nuclease A gene ( SN ) . Linker sequence added to the primer design , using the touch down PCR method to realize E and SN gene series ( E - 15aalinker - SN ) . E gene E-15aalinker-SN gene insert Thermostat the expression vector pBV220 , and building temperature-controlled single- gene expression system pBV-E and temperature control gene expression system pBV - E - 15aalinker -SN . To pBV-E-15aalinker-SN plasmid as a template the thermostat double Lysis Box (DLS) was amplified by PCR method to construct the recombinant cloning vector pMD- WK . Digestion DLS, insert it into the E. coli-App shuttle vector ( PMC- Express ) build the thermostat double Lysis carrier of pMC- WK . Cracking tests showed that the cracking superior of the E - 15aalinker - SN E, showed cleavage rapid and thorough , cell cracking rate of up to 99.999% ; by PCR amplification of the DLS has biological activity , indicating that the DLS successful amplification . By bonding conversion the pMC-WK import pleuropneumonia Actinobacillus (Actinobacillus pleuropneumoniae, App), and successfully prepared App bacterial ghost . Pasteurella multocida (Pasteurella multocida, Pm) genome as a template , and amplified by PCR outer membrane protein ompH gene . Eukaryotic expression vector pCDNA3.1 () is the skeleton construct pCDNA - of ompH is . Its as a DNA vaccine , by soaking the the pCDNA-ompH plasmid Loading App bacteria shadow . Immune trials show that the to : App bacterial ghost pleuropneumoniae Actinobacillus pig Pasteurella multocida infection has a protective effect , the protection rate of up to 100% load the pCDNA-ompH vaccine .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Pig
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