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Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)Nghiên cứu đặc tính của Chitinase tự nhiên và biểu hiện Chitinase tái tổ hợp từ chủng nấm Lecanicillium Lecanii (Luận văn thạc sĩ)
N VÀ LECANICILLIUM LECANII T i Trung tâm H , 2015 - http://www.lrc-tnu.edu.vn/ N VÀ LECANICILLIUM LECANII Chuyên ngành: 62 42 01 16 : T i Trung tâm H , 2015 - http://www.lrc-tnu.edu.vn/ i , , - ,m M i Trung tâm H - http://www.lrc-tnu.edu.vn/ ii , TS Ng Tôi xin chân thành s E cho q trình Lecanicillium thơn -2013 , Khoa Sinh -K q G - i Trung tâm H - http://www.lrc-tnu.edu.vn/ iii Trang i ii iii vii HÌNH ix xii Lecanicillium lecanii 1.2 Chitinase 12 14 16 i Trung tâm H - http://www.lrc-tnu.edu.vn/ iv L lecanii chitinase 19 1.3 19 1.3.2 Trong 1.3 công 24 1.4 1.4.1 1.4.2 23 óa chitinase 25 25 29 33 33 2.1.2 33 33 33 34 2.1.4 Du 34 33 33 33 35 2.2.3 2.2.4 Các p 2.2.7 3.1 S 42 47 lý 49 50 , i Trung tâm H - http://www.lrc-tnu.edu.vn/ v L lecanii 50 3.1.1 L lecanii 50 3.1.2 L lecanii 50 53 L Lecanii 43H 61 61 L lecanii 3.2.2 43H 3.3 Nhân dòng gen mã hóa chitina 64 L lecanii 43H 67 P pastoris X33 71 n gen Chit 71 P pastoris 72 77 P pastoris X33 79 tính L lecanii 85 85 87 88 89 L lecanii 43H i Trung tâm H - 90 http://www.lrc-tnu.edu.vn/ vi 93 93 93 94 95 114 i Trung tâm H - http://www.lrc-tnu.edu.vn/ vii Ti ng Anh Ti ng Vi t BLAST Basic local alignment search tool Ph n m m so sánh trình t bp Base pair C cDNA Complement DNA DNA b sung Chit Gene encoding chitinase Gen mã hóa chitinase CMC Cacboxyl methyl cellulose Cacboxyl methyl cellulose DEPC Diethylpyrocarbonate Diethylpyrocarbonate DNA Deoxyribonucleic acid Axit deoxyribonucleic DNase Deoxyribonuclease Enzyme th y phân DNA dNTPs -Deoxynucleoside - Các nucleotide triphosphate Control IPTG i ch ng Isopropyl-beta-D- Isopropyl-beta-D- thiogalactopyranoside thiogalactopyranoside EDTA Ethylenediamine tetraacetic acid Axit ethylenediamine tetraacetic EtBr Ethidium bromide Ethidium bromide kb Kilo base Kilo base kDa Kilo Dalton Kilo Dalton M Marker Thang chu n OD Optical density M PCR Polymerase chain reaction Ph n ng khu RNA Ribonucleic acid Axit ribonucleic i Trung tâm H - quang i gen http://www.lrc-tnu.edu.vn/ viii RNase Ribonuclease Enzyme th y phân RNA RT-PCR Reverse transcription polymerase Ph n ng khu i gen chain reaction rChit Recombinante chitinase SDS- Sodium dodecyl sulfate PAGE Polyacrylamide gel Chitinase tái t h p n di bi n tính protein gel polyacrylamide electrophoresis Taq Thermus aquaticus Thermus aquaticus TBE Tris boric acid EDTA Tris boric acid EDTA TE Tris EDTA Tris EDTA TEMED N,N,N ,N - N,N,N ,N - Tetramethylethylenediamine Tetramethylethylenediamine v/v Volume/volume Th tích/th tích w/v Weight/volume Kh i Trung tâm H - ng/th tích http://www.lrc-tnu.edu.vn/ 107 44 Ghanem K M., Al-Garni S M., Al-Makishah N H (2010), "Statistical optimization of cultural conditions for chitinase production from fish scales waste by Aspergillus terreus", Afr J Biotechnol., 9, pp 5135-5146 45 Gomaa E Z (2012), "Chitinase production by Bacillus thuringiensis and Bacillus licheniformis: their potential in antifungal biocontrol", J Microbiol., 50, pp 103-11 46 Gooday G W., Humphreys A M., McIntosh W H (1986), "Roles of chitinases in fungal growth In: RAA Muzzarelli, C Jeuniaux, GW Gooday (eds): Chitin in nature and technology", Plenum Press, New York, pp 83 -91 47 Gopalakrishnan C (1989), "Susceptibility of cabbage diamondback moth Plutella xylostella L to the entomofungal pathogen Verticillium lecanii (Zimmerman), Viegas", Current Science, 58, pp 1256-1257 48 Hall R A (1981), "The fungus Verticillium lecanii as a microbial insecticide againt aphids and scales In: Microbial control of pests and plant disease 19701980 (H.D Burges, ed)", Academic Press London, pp 483-498 49 Hall R A (1981), "A new insecticide against greenhouse aphids and whitefly: the fungus Verticillium lecanii", Ohio Florists Assoc Bull, 626, pp 3-4 50 Hamsah N B (2009), "Optimization of inducers on expresion of recombinant chitinase in Escherichia coli using response surface methodlogy", Faculty of Chemical & Natural Resources Engineering Universiti Malaysia Pahang, pp 1-24 51 Harma G E., Hayes C K., Lorito M., Broadway R M., Di pietro A., Peterbauer C., Tronsmo A (1993), "Chitinolytic enzymes of Trichoderma harzinum: purification of chitobiosidase and endochitinase", Phytopathology, 83, pp 313-318 i Trung tâm H - 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http://www.lrc-tnu.edu.vn/ 120 Ammonium persulfate 10% Dung 100 ml ethanol 95% (v/v), 350 mg serva blue G, 200 ml phosphoric acid 88% (v/v) 425 ml H2O, 15 ml ethanol 95% (v/v), 30 ml working -HCl 1,5M, pH 8,8 -HCl 0,5M, pH 6,8 Acrylamide 30%, bis-acrylamide 0,8% SDS 10% DNS 0,53%, NaOH 0,99%, KNaC4H4O6 15,3%, Na2S2O5 0,415%, phenol 0,38% (v/v) Tris-HCl 100 mM, EDTA 10 mM, pH 8,0 NaOH 0,2 N, SDS 1% K 28,5% Ethidium bromide 0,1 g/ml Coomassie brilliant blue 0,1%, methanol 30% PAGE (v/v), acid acetic 10% Methanol 30% (v/v), acid acetic 10% (v/v) i Trung tâm H - http://www.lrc-tnu.edu.vn/ 121 Tris-HCl 20 mM, glycine 192 mM, SDS 0,1%, pH 8,8 Brommophenol blue 0,05%, glycerol 50% pha -HCl M, pH 6,8 Tris-HCl 10 mM, EDTA mM, pH EDTA 20 mM, Tris-borate 900 mM, pH Brommophenol blue 0,09%, xylene xyanol 0,09%, glycerol 60% Ethanol 70% (v/v) i Trung tâm H - http://www.lrc-tnu.edu.vn/ ... http://www.lrc-tnu.edu.vn/ 1.1 Lecanicillium lecanii Lecanicillium Lecanicillium Lecanicillium lecanii, L attenuatum, L longisporum, L muscarium L nodulosum L lecanii Verticillium lecanii Ngày nay, L lecanii Cordyceps... [124] Lecanicillium [22], Lecanicillium [47], [49], [60], [93] [95] L lecanii là: L lecanii L lecanii sinh Lecanicillium C L lecanii i Trung tâm H - http://www.lrc-tnu.edu.vn/ L lecanii 1.2 Chitinase. .. enzyme (chitinase, protease -acetyl -1,4-glucosid C4 - chitin i Trung tâm H - http://www.lrc-tnu.edu.vn/ : Lecanicillium lecanii L lecanii Pichia pastoris chitinase X33 L lecanii cao ; L lecanii