Surface protein of streptococcus suis type-2, preparation method thereof and application
A technology of Streptococcus suis and surface protein, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve problems such as difficult to achieve ideal effects, and achieve the effect of broad market prospects.
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Embodiment 1
[0016] Example 1 Expression, purification and antibody preparation of Streptococcus suis type 2 surface protein SSU98_1675
[0017] 1. Materials and Methods
[0018] 1.1 Strains and plasmids
[0019] Streptococcus suis type 2 98HAH12 strain [Chen C, Tang J, Dong W, Wang C, Feng Y, et al(2007) A Glimpse of Streptococcal Toxic Shock Syndrome from Comparative Genomics of S.suis 2 Chinese Isolates.PLoS ONE 2(3) : e315]; Escherichia coli DH5a, BL21, are international standard strains; expression vector pET32a (+) is a product of Novagen.
[0020] 1.2 Enzymes and Reagents
[0021] EcoRI, XhoI and other restriction endonucleases, as well as Taq enzymes, dNTPs, DL15000 molecular weight standards and other PCR reagents are all products of Bao Biological Engineering (Dalian) Co., Ltd.; DNA recovery kits are products of Tianwei Company; nickel affinity chromatography column It is a product of GE Company; the plasmid extraction kit is a product of V-gene Company; Freund's complete adju...
Embodiment 2
[0048] Example 2 Immunogenicity of Streptococcus suis type 2 surface protein SSU98_1675 and antibody-mediated opsonophagocytosis test
[0049] 1. Material method: material is the same as embodiment one.
[0050] 1.1 Preparation of recombinant Streptococcus suis type 2 surface protein SSU98_1675 and polyclonal antibody: see Example 1.
[0051] 1.2 Opsonophagocytosis mediated by SSU98_1675 antibody
[0052] Scrape a single colony of Streptococcus suis type 2 from the blood plate, insert into THB medium, 37°C, CO 2 After culturing in an incubator for 8 hours, transfer to fresh THB medium and culture for 8 hours to logarithmic growth phase. Take 1ml bacterial solution (about 1×10 8 CFU / ml) 13,000rpm, collect bacteria for 1min, wash with PBS and resuspend, adjust to 10 6 CFU / ml, smear blood plate, recorded as the initial bacterial count. Take 50μl (≈10 6 CFU / ml) bacteria solution, add 100 μl antibody, 15 minutes at 37°C and then put on ice for 15 minutes; add 350 μl healthy h...
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