Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)Nghiên cứu tạo kháng nguyên tái tổ hợp STAPHYLOCOCCAL ENTEROTOXIN B ( SEN) dạng không độc từ đoạn GEN SEB tự nhiên làm nguyên liệu phục vụ chế tạo que thử phát hiện nhanh độc tố SEB (Luận văn thạc sĩ)
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PGS TS Hàn lâm Khoa giúp tơi hồn thành Tơi xin c chân thành chun môn Tôi th 15: phylococcal enterotoxin B (SEB) TS 2013 2015 , tơi xin tháng Hồi Thu i Trung tâm H http://www.lrc-tnu.edu.vn/ Trang i iii .v Staphylococcus aureus) 1.1.2 T i Vi t Nam Staphylococcus aureus .11 15 .15 16 17 17 21 23 1.4.1 .23 24 25 i Trung tâm H http://www.lrc-tnu.edu.vn/ .26 .26 26 26 28 28 28 .28 29 PCR) .30 30 .31 -primer 31 32 33 33 enzym .34 34 35 .35 .36 37 38 i Trung tâm H http://www.lrc-tnu.edu.vn/ 38 40 42 S aureus nhân dòng gen seb 42 S aureus .42 3.1.2 Nhân dòng gen seb 42 .44 seb 45 45 .47 seb (b+) 48 .48 3.3.2 G .49 3.4 T gen seb E coli BL21 50 oli BL21 50 51 .55 Bradford 57 58 58 a protein wtSEB mtSEB LD50 lô .58 i Trung tâm H a protein wtSEB mtSEB LD50 60 a protein wtSEB mtSEB 10xLD50 61 http://www.lrc-tnu.edu.vn/ K T LU N 63 63 64 75 i Trung tâm H http://www.lrc-tnu.edu.vn/ µl Microliter APS Ammonium persulphate bp Base pair BSA Bovine serum albumin dH2O DNA Deoxyribonucleic acid DNase Deoxyribonuclease dNTP Deoxyribonucleotide EDTA Ethylene Diamine Tetraacetace Axit IPTG K16 -D-1-thiogalactopyranosid Kb Kilobase kDa Kilo Dalton LB Luria - Bertani LD50 Lethal dose, 50% ml Milliliter mtSEB Mutant SEB OD Optical density PBS Phosphate buffer saline PCR Polymerase Chain Reaction RNA Ribonucleic Acid S aureus Staphylococcus aureus SDS-PAGE Sodium dodecyl sulfate polyacrylamide gel electrophoresis i K16 SEB Staphylococcal enterotoxin B SEs Staphylococcal enterotoxins TAE Tris-acetate-EDTA Taq Thermus aquaticus TE Tris EDTA TEMED N,N,N ,N -Tetramethylethylenediamine v/p Vòng/phút v/v volume/volume w/v Weight/volume wtSEB Wild type SEB ii Hình 3.20 LD50 sau ngày theo dõi 3.6.3 a protein wtSEB mtSEB tiêm 10xLD50 10xLD50 10xLD50 (= wtSEB (lơ 4) (hình 3.21 lơ tiêm mtSEB xLD50 (= 25 µg/g) 3.5 wtSEB, mtSEB tiêm10xLD50 (%) Lô (tiêm ActD M2 = 10xLD50) 10/10 100 Lô (tiêm ActD M4 = 10xLD50) 0/10 K16 61 Hình 3.21 m 10xLD50 sau ngày theo dõi sau: xLD50; 3xLD50 10xLD50) k protein wtSEB tiêm 1xLD50 xLD50 10xLD50 K16 62 K T LU N seb S aureus seb amino acid Histidine thành Tyrosine gen seb E coli BL21 o Tinh NTA bình mg/ml 10xLD50 (25 µg/g) protein wtSEB tiêm 1xLD50 (2,5 µg/g) xLD50 (7,5 µg/g) 10xLD50 (25 µg/g) K16 63 T im Dung (2005) Chí Minh, Thông ] , Thông tin khoa 179 trang Agrawal R, Singh PK, Sharma SK, Kamboj DV, Goel AK, Singh L (2012) Highly expressed recombinant SEB for antibody production and development of immunodetection system Indian J Microbiol 52: 191 196 Staphylococcus aureus isolated from bovine raw milk and Minas frescal cheese in Brazil J Food Prot 73(12): 2225 2231 Balaban N, Rasooly A (2000) Staphylococcal enterotoxins Int J Food Microbiol 61: 1-10 Barber MA (1914) Milk poisoning due to a type of Staphylococcus albusoccurring in the udder of a healthy cow Philipp J Sci 9: 515 519 K16 64 10 Barik S (1993) Site-directed mutagenesis by double polymerase chain reaction Methods Mol Biol 15: 277-286 11 Bayles KW, Iandolo JJ (1989) Genetic and molecular 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Zhang T, Feng Y, Lu X, Lan W, Wang J, Wu H, Cao C, Wang X (2011) Highly efficient production of soluble proteins from insoluble inclusion bodies by a two-step-denaturing and refolding method PLoS ONE 6: e22981 K16 74 hùy seb 534 nucleotide 12(1): 173-180 K16 75 ... thi n nguyên li u quan tr ng ch t o que th phát hi n nhanh SEB ph c v nhu c u hi n c lý trên, chúng tài SEB seb SEB nhanh seb S aureus gen seb amino acid Histidine thành Tyrosine seb seb ánh... 30 ô gel tách seb 36 S aureus 57 wtSEB, mtSEB tiêm 1xLD50 wtSEB, mtSEB tiêm 3xLD50 tiêm 10xLD50 K16 45 wtSEB, mtSEB 59 60 61 v T c u vàng (Staphylococcus aureus aureus) m t nh ng nguyên nhân gây... dòng gen seb 42 S aureus .42 3.1.2 Nhân dòng gen seb 42 .44 seb 45 45 .47 seb (b+ ) 48 .48 3.3.2 G .49 3.4 T gen seb E coli BL21