Method for constructing swine escherichia coli and haemophilus parasuis attenuated fusion strain
A technology of Haemophilus suis and Escherichia coli, which is applied in the preparation of hybrid cells to achieve the effects of reducing virulence, improving immune effect, and increasing unit antigen content
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Embodiment 1
[0028] Example 1 Screening of Escherichia coli E.coli and Haemophilus parasuis HPS fusion parental drug-resistant strains
[0029] 1 material description
[0030] 1.1 Strains
[0031] The identified serotype is O 107 Escherichia coli E.coli and Haemophilus parasuis HPS are preserved by our laboratory.
[0032] 1.2 Main reagents
[0033] Various drug-sensitive discs were purchased from Hangzhou Tianhe Microbiological Reagent Co., Ltd.; antibiotics rifampicin and amikacin sulfate were purchased from Dalian Meilun Biotechnology Co., Ltd.
[0034] 1.3 Main medium
[0035] plain broth;
[0036] Ordinary agar medium;
[0037] TSB liquid medium;
[0038] TSA medium.
[0039] 2 Methods and results
[0040]2.1 Drug susceptibility test by disc diffusion method
[0041] Recover Haemophilus parasuis HPS on a TSA agar plate at 37°C for 18-24 hours, and Escherichia coli on an ordinary agar plate at 37°C for 12-18 hours, and pick a single Haemophilus parasuis HPS and E. coli E.col...
Embodiment 2
[0057] Example 2 Construction and screening of pig Escherichia coli E.coli and Haemophilus parasuis HPS fusion strain
[0058] 1 material description
[0059] 1.1 Strains
[0060] The identified serotype is O 107 Escherichia coli E.coli and Haemophilus parasuis HPS are preserved by our laboratory.
[0061] 1.2 Main reagents
[0062] Antibiotics rifampicin and amikacin sulfate were purchased from Dalian Meilun Biotechnology Co., Ltd.; lysozyme (Lysozyme, 40000U / mg) was purchased from Amresco Company; deoxyribonuclease I (DNaseI, 2000U / mg) was purchased from Roche Company The fusion agent polyethylene glycol PEG4000 was purchased from Merck; the positive serum of Haemophilus parasuis was purchased from Guangdong Dahuanong Animal Health Products Co., Ltd.; the positive serum of E. Pure reagents.
[0063] 1.3 Main solution
[0064] Tris buffer: 0.01mol / L Tris, pH7.0
[0065] Hypertonic Tris buffer: 0.01mol / L Tris, 0.5mol / L sucrose, 0.01mol / L CaCl 2 , pH7.0
[0066] SMM bu...
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