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Construction and preliminary application of the expression of two different outer membrane protein of avian Escherichia coli fusion strains

Author: YuYanRong
Tutor: ZhangYanMing
School: Northwest University of Science and Technology
Course: Preventive Veterinary Medicine
Keywords: Avian E. coli Outer membrane protein Protoplast fusion Vaccine development
CLC: S852.61
Type: Master's thesis
Year: 2001
Downloads: 139
Quote: 5
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Abstract


Major serotypes isolated from the Shaanxi Province chicken regional disease dead chickens and chicken embryos pathogenic avian E. coli outer membrane protein (Omp) extraction using ultrasonic lysis and N- dodecane sarcosine , OMP typing results by SDS-PAGE , the 18 strains of the the three kinds Omp type , wherein four O1, 4 strains of O2 , 4 strains O78 and 3 O89 has two the Omp -type ( omp1 2 type) the 3 O75 the three the Omp type ( omp1 2,3 type) , between the different serotypes common Omp type , and the the same serotype Omp type can be different . Lysozyme lysis cell wall with different concentrations and different time preparing protoplasts in hypertonic plates Regeneration Culture Select protoplast formation and regeneration of the best conditions for the enzyme the concentration 6mg/mL role time 25min ; susceptibility test based on , to combined the Omp analysis , select cross-resistance and a different type of Omp strains , PEG-induced protoplast fusion , screening in hypertonic plate with double anti fusant different generation fusion strains were Gram 's staining , biochemical identification , serotype identification and Omp analysis , the results showed that the staining, and biochemical characteristics are in line with characteristics of E. coli ; the 75% fusants O antigen agglutination two parents , 25% agglutination one parent O antigen ; outer membrane proteins of the performance of one or two parents with type , but has a different degree of enhancement . The stability test to prove that it is genetically stable integration of the sub . Made the seedlings bacteria preparation E.coli alhydrogel seedlings immunized animals with prepared fusant as antibody levels measured and challenged the experiment , and the unit price Miao comparative assessment of the integration of the immune effects of strain vaccine . The results show that : the fusion strains vaccine antibody levels Antibody Dynamic Price seedlings is basically the same , but the fusion strain vaccine protection rate of the different serotypes of E.coli hybrid challenged Price seedlings and control improved from 13% to 33 % and from 60% to 66% , indicating that the expression of two different outer membrane protein the fusion strain vaccine antigen spectrum broader and stronger protection .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Pathogenic bacteria
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