Surface protein of streptococcus suis 2-type and preparation method and application thereof
A surface protein, Streptococcus suis technology, applied in the field of bacterial proteins, can solve problems such as difficulties in identifying immunogenic proteins, false positive reactions, and inability to achieve high-throughput screening
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Embodiment 1
[0021] Example 1 Expression and purification of Streptococcus suis type 2 surface protein 0186, immunogenicity and antibody-mediated opsonophagocytosis test
[0022] 1 Materials and methods
[0023] 1.1 Strains and plasmids
[0024] 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.
[0025] 1.2 Enzymes and Reagents
[0026] 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-gen...
Embodiment 2 86
[0066] The preparation process of embodiment two 186 colloidal gold detection test strips
[0067] 1. Antibody Processing
[0068] The immune serum obtained by immunizing rats with 186 and New Zealand rabbits with Streptococcus suis was purified by caprylic acid-ammonium sulfate precipitation or protein A column, dialyzed overnight with 0.01M PBS, centrifuged at 10,000r / min for 30min, and collected the supernatant to measure the concentration. use.
[0069] 2. Preparation of colloidal gold (sodium citrate reduction method)
[0070] HAuCl 4 First make a 1% aqueous solution, take 100ml of pure water and heat it to boiling. Add 1ml of 1% HAuCl while stirring 4 and 1.5 ml of trisodium citrate in water. Continue to heat and boil for about 15 minutes. After adding the liquid, it is observed that the color turns black first, and then slowly turns into wine red. After seeing the wine red, continue heating for 3-5 minutes and then stop heating. The obtained is colloidal gold with a...
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