Antigen simulating epipeptide of A type parachicken blood phili bacillus and its use
A technology of Haemophilus gallinarum and mimetic epitope, which is applied in the field of short peptides simulating the antigenic epitope of Haemophilus paragallinarum to achieve the effects of simplifying the vaccine production process, reducing the chemical residues of vaccines and reducing the influence of HPG endotoxin
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Embodiment 1
[0029] Example 1: Screening and sequence analysis of Haemophilus paragallinarum antigen mimotope
[0030] Experimental Materials
[0031] 1 Phage Random Peptide Library Kit
[0032] Linear 12-peptide library displayed on phage surface capsid protein III (Ph.D.-12 TM Phage Peptide Library PeptideLibrary Kit) kit was purchased from New England Biolabs, USA. The titer of the peptide library was 4×10 12 pfu / ml, random diversity 1.9×10 9 ; The genotype of the recipient strain E.coli ER2537 is F'lac q Δ(lacZ)M15proA + B + fhuA2 supE thiΔ(lac-proAB)Δ(hsdMS-mcrB)5(r k - m k - McrBC - ). DNA automatic sequencing primer (-96gIII sequencing primer): 5'-CCCTCATAGTTAGCGTAACG-3'
[0033] Polyantisera of Haemophilus paragallinarum Type 2A and Type C and Chickens Artificially Infected with HPg A and HPg C
[0034] Haemophilus paragallinarum type A: 0083 strains; Haemophilus paragallinarum type C: 668 strains and Modesto strain are all preserved by the Animal Husbandry and Veter...
Embodiment 2
[0114] Example 2: Application of Mimotope Fusion Peptides in Vaccines
[0115] Experimental Materials
[0116] Host bacteria: GI826(F - ,lacI q ,ampC∷P trp cI, ΔfliC, ΔmotB, eda::Tn10), product of Invitrogen Company; plasmid: pFliTrx product of Invitrogen Company;
[0117] main reagent
[0118] Restriction endonuclease: ApaI, product of Xho I TaKaRa company;
[0119] T4DNA ligase: TaKaRa company product;
[0120] Taq polymerase: product of TaKaRa Company.
[0121] Related media
[0122] 10XM9 Salt: Na 2 HPO 4 60g, KH 2 PO 4 30g, NaCl 5g, NH 4 Dissolve 10g of Cl in 900ml of deionized water, and adjust the pH to 7.4 with 10M NaOH;
[0123] IMC medium: containing 1×M9 salt solution, 0.2% casein acid hydrolyzate, 0.5% glucose, 1mmol / L MgCl 2 ;
[0124] RM medium: 1XM9 salts, 2% tyrosine, 1% glycerol, 1mMMgCl 2 ;
[0125] SOC medium: tryptone 20g, yeast extract 5g, NaCl 0.5g, 2.5mmol / L KCl, 10mmol / LMgCl 2 , 20mmol / L glucose.
[0126] equipment
[0127] BIO-Rad G...
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