Listonella anguillarum subunit vaccine antigenic protein and application thereof
A technology of Listonia anguillaris and subunit vaccines, applied in the field of immunology, to achieve the effects of increasing immune synergy, broadening the vaccine protection spectrum, and simple preparation methods
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Embodiment 1
[0021] According to the nucleotide sequence of the Listonia anguillarum T6SS matrix protein gene lahcp, and the restriction map design amplification primer of vector pETDuet (purchased from Novagen):
[0022] lahcp-for-EcoRI:5'-GGC GAATTC ATGCCAACTGCATG-3' (underlined base sequence is the recognition site of restriction endonuclease EcoRI);
[0023] lahcp-rev-SalI:5'-ATCG GTC GAC TTACGCTTCAACAG3' (the base sequence underlined is the recognition site of restriction endonuclease SalI).
[0024] The PCR amplification reaction was carried out using the Genomic DNA of Listonia anguillarum as a template (the PCR instrument was purchased from Takara Company, model TP600), and the reaction system and reaction parameters were as follows: (Reagents should be expressed in Chinese as much as possible)
[0025] PCR reaction system (25μL)
[0026] 10×PCR buffer 2.5μL
[0027] dNTP mixture 0.5μL (each 0.2mM)
[0028] MgCl 2 2.0μL (1.5mM)
[0029] Primer lahcp-for-Ec...
Embodiment 2
[0106] Mix the antigenic protein of the Listeria anguillaris subunit vaccine with complete Freund's adjuvant at a ratio of 1:1 by volume, and then intraperitoneally inject it into the turbot with an inoculation dose of 2 μg protein / g fish body weight .
Embodiment 3
[0108] Mix the antigenic protein of the Listeria anguli subunit vaccine with complete Freund's adjuvant in a ratio of 1:1 by volume, and then intraperitoneally inject it into the blue eel dragon with an inoculation dose of 20 μg protein / g fish body weight .
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