Phân lập và phân tích genome thực khuẩn thể kháng vi khuẩn gây bệnh quan trọng trên cá tra

I H C QU C GIA TP.HCM I H C BÁCH KHOA NGUY N LÊ HOÀI NHÂN PHÂN L P VÀ PHÂN TÍCH GENOME TH C KHU N TH KHÁNG VI KHU N GÂY B NH QUAN TR NG TRÊN CÁ TRA Chuyên ngành: Công ngh sinh h c Mã s : 60420201 LU TP H c hoàn thành t i: i H c Bách Khoa Cán b ng d n khoa h c 1: PGS.TS Lê Phi Nga Cán b ng d n khoa h c 2: TS Hoàng Anh Hoàng HCM Cán b ch m nh n xét 1: TS Nguy Cán b ch m nh n xét 2: TS Phan Th Huy n Lu cb ov t iH Thành ph n H m: PGS.TS Nguy n Ti n Th ng Ch t ch TS Hu nh Ng c Oanh T TS Nguy Ph n bi n TS Phan Th Huy n Ph n bi n PGS.TS Lê Phi Nga U viên Xác nh n c a Ch t ch H ngành sau lu CH T CH H ng Khoa qu n lý chuyên c s a ch a (n u có) NG NG KHOA K THU T HĨA H C I H C QU C GIA TP.HCM C NG HÒA XÃ H I CH I H C BÁCH KHOA T NAM c l p - T - H nh phúc NHI M V LU nh : 60420201 I TÀI Phân l p phân tích genome th c khu n th kháng vi khu n gây b nh quan tr ng cá tra II NHI M V VÀ N I DUNG Phân l p kh c tính vi sinh c a th c khu n th Edwardsiella ictaluri gây b nh cá tra t m u bùn Phân l p nh ph xâm nhi m Kh o sát ho t tính xâm nhi m m genome c a th c khu n th Phân tích genome th c khu n th kháng Aeromonas hydrophila phân l p Vi i genome c a PVN02 phân l p Vi 2020 Phân tích genome th c khu n th kháng Edwardsiella ictaluri phân l p Vi t Nam hi III NGÀY GIAO NHI M V : 22/02/2021 IV NGÀY HOÀN THÀNH NHI M V : 13/06/2021 V CÁN B NG D N: PGS.TS Lê Phi Nga, TS Hoàng Anh Hoàng CÁN B PGS.TS Lê Phi Nga CH NHI M B PGS.TS Lê Th Th y Tiên O NG D N TS Hoàng Anh Hồng NG KHOA K THU T HĨA H C Lu L IC Trên th c t khơng có s thành công mà không g n li n v i nh ng h tr , giúp dù hay nhi u, dù tr c ti p hay gián ti p c b uh ct p tâm gi ng Cao h c a quý Th c nh t, Em xin g v n quý Th y, Cô Khoa Công ngh i h c Bách Khoa Thành Ph H tâm huy t c gian h c t p t c r t nhi u s quan n bè V i lòng bi Hóa H c i khác Trong su t th i gian t truy i tri th c t v n ki n th c quý báu cho chúng em su t th i ng Sau th i gian h c t p nghiên c p phân tích genome th c khu n th kháng vi khu n gây b nh quan tr Hoàn thành khóa lu n c bày t l i c i Cô PGS TS Lê Phi Nga, Th y TS Hồng Anh Hồng, Cơ Th y - ng d n ng th i c y Cô giáo Em Xin c c bi t ch Tr n Th Thanh Xuân anh T em r t nhi u M u c g ng h c t p nghiên c u su t th i gian qua, song th i gian có h n, nên lu i nh ng thi u sót Em r t mong nh n c s góp ý c a Th y Cơ giáo nh c hoàn thi HVTH: Nguy n Lê Hồi Nhân lu a Trang i Lu TĨM T T B nh gan th n m b nh xu t huy t gây b i vi khu n Edwardsiella ictaluri Aeromonas hydrophila hai nh ng b nh quan tr ng cá tra (Pangasianodon hypophthalmus) t i ao nuôi thu d ng b ng b ng Sơng C u Long Nh m góp ph p th c khu n th , n phân l c 13 th c khu n th xâm nhi m Edwardsiella ictaluri t m u bùn K t qu kh o sát ho t tính xâm nhi m Edwardsiella ictaluri c th c hi n th c khu n th phage M14N có chu k lây nhi m h s nhân t t nh t 100 phút, 160.52 ± 3.12 K t qu nh ph xâm nhi m kh c th c khu n th M18N có xu t hi n khu v c ly gi i c dòng vi khu n có ph ký ch r ng nh t Edwardsiella ictaluri E1, Edwardsiella ictaluri E3-1, Edwardsiella ictaluri E4-2, Edwardsiella ictaluri E5-3, Edwardsiella ictaluri E6-4 Trong nghiên c u này, 13 th c khu n th phân l th kh c tách chi t genome Và nghiên c c phân ch d d li phân tích genome t th c khu n th A hydrophila m m E ictaluri c phân l p tách chi m t nhóm nghiên c u c i ih c b gen c a th c khu n th PVN03, PVN04, PVN05, PVN06, PVN07 PVN08 l 50.698 bp, 51.720 bp, 51.670 bp, 43.645 bp, 43.235 bp 42.439bp S t cm (orf) c a th c khu n th l t 64 (orf), 65 (orf), 66 (orf), 69 (orf), 60 (orf) 66 (orf) T t c tRNA, rRNA ho c sRNA, gen kháng thu c kháng sinh, y u t gen tích h cl u khơng có c t c phát hi n sáu b gen c a phage Bên c c phân tích phát sinh lồi nhóm th c khu n th PVN03, PVN04, PVN05 v i PVN02 c a nhóm nghiên c u Tu Q Vinh c ng s (2020) c phân l p t i Vi Aeromonas hydrophila o thành m t nhóm riêng bi t v i th c khu n th d li u y nhóm th c khu n th PVN06, PVN07, PVN08 v i Pei21 t o thành m t nhóm v i v i th c khu n th HVTH: Nguy n Lê Hoài Nhân Trang ii Lu ABSTRACT Hepatobiliary disease and hemorrhagic disease caused by Edwardsiella ictaluri and Aeromonas hydrophila are two important diseases of catfish (Pangasianodon hypophthalmus) farmed in the Mekong Delta In order to contribute to the diversity of the phage collection, in this study, I have preliminarily isolated 13 Edwardsiella ictaluri bacteriophages from the sludge sample The survey results of Edwardsiella ictaluri infectivity were performed on phages, including the M14N phage line with the best infectivity cycle and multiplier of 70 minutes, 160.52 ± 3.12 The results of infection spectrum determination were that the phage M18N appeared in the lysis region on all bacterial strains and had the widest host spectrum Edwardsiella ictaluri E1, Edwardsiella ictaluri E3-1, Edwardsiella ictaluri E4-2, Edwardsiella ictaluri E5-3, Edwardsiella ictaluri E6-4 In this study, 13 isolated bacteriophages have not been subjected to the genome analysis step because there is not enough time to investigate all the microbiological characteristics as well as the genome extraction step And the study used the database to analyze the genome from new A hydrophila phages such as PVN03, PVN04, PVN05 and new E ictaluri such as PVN06, PVN07, PVN08 that had been isolated and extracted genomes by a research group of Ho Chi Minh City University of Technology The genome sizes of phages PVN03, PVN04, PVN05, PVN06, PVN07 and PVN08 were 50,698 bp, 51,720 bp, 51,670 bp, 43,645 bp, 43,235 bp and 42,439bp, respectively The open reading frame (orf) of phages PVN03, PVN04, PVN05, PVN06, PVN07 and PVN08 were also 64 (orf), 65 (orf), 66 (orf), 69 (orf), 60 (orf) and 66 (orf) All were free of tRNA, rRNA or sRNA, antibiotic resistance genes, virulence factors, toxin genes and integrative genes detected in the six phage genomes Besides, the analysis of phylogenetic tree groups of phage groups PVN03, PVN04, PVN05 together with PVN02 of the research group Tu Q Vinh et al (2020) isolated in Vietnam has formed a separate group with other Aeromonas hydrophila bacteriophages in the database In addition, the analysis of the phylogenetic tree also showed that the bacteriophages PVN06, PVN07, PVN08 together with Pei21 form a group with the bacteriophages previously present on NCBI HVTH: Nguy n Lê Hoài Nhân Trang iii Lu L Phân l p phân tích genome th c khu n th kháng vi khu n gây b nh quan tr ng cá tra v is u c a ng d n c a PGS.TS Lê Phi Nga TS Hoàng Anh Hoàng Các s li u k t qu trình bày lu i khác cơng b b t k cơng trình khác TÁC GI LU NGUY N LÊ HOÀI NHÂN HVTH: Nguy n Lê Hoài Nhân Trang iv Lu M CL C L IC i TÓM T T ii L iv M C L C v DANH M C CH VI T T T viii DANH M C CÁC HÌNH ix DANH M C CÁC B NG xi M U T NG QUAN TÀI LI U 1.1 Giá tr kinh t ngành nuôi cá tra t i Vi t Nam .2 1.2 Tình hình d ch b nh gan th n m b nh xu t huy t cá tra b t c p s d ng kháng sinh .2 1.2.1 Tình hình d ch b nh gan th n m cá tra 1.2.2 Tình hình d ch b nh xu t huy t cá tra 1.2.3 B t c p s d 1.3 u tr d ch b nh th y s n .4 T ng quan v th c khu n th li u pháp th c khu n th 1.3.1 T ng quan v th c khu n th 1.3.2 Li u pháp th c khu n th 1.4 Cơng ngh gi i trình t gene 1.4.1 Gi i trình t gen th h m i b ng Illumina 1.4.2 L p ráp gi i 12 1.5 Tình hình nghiên c c 14 1.5.1 c 14 1.5.2 c 14 1.6 M c tiêu n i dung nghiên c u c tài 17 1.6.1 M c tiêu 17 1.6.2 N i dung nghiên c u .17 2.1 V T LI .19 V t li u 19 HVTH: Nguy n Lê Hoài Nhân Trang v Lu 2.1.1 Th m nghiên c u 19 2.1.2 2.1.3 ng nghiên c u 19 Hóa ch 2.2 ng, thi t b d ng c 19 19 Thí nghi m 1: Phân l p kh c tính vi sinh c a th c khu n th i kháng vi khu n Edwardsilla ictaluri gây b nh cá tra t m u bùn 19 2.2.1 t ng quát 20 2.2.2 Thu x lý m u 20 2.2.3 Phân l p th c khu n th .21 2.2.4 Tinh s ch t o stock th c khu n th 22 2.2.5 Kh o sát ho t tính xâm nhi m c a th c khu n th p 23 2.2.6 nh ph xâm nhi m c a th c khu n th 25 Thí nghi m genome c a th c khu n th 26 2.2.1 Ki m tra x lý d li u thô 26 2.2.2 L p ráp h gen 27 2.2.3 Chú gi i c u trúc 27 2.2.4 Chú gi i ch 2.2.5 Phân tích phát sinh lồi 29 28 K T QU VÀ TH O LU N 30 3.1 Thí nghi m 1: Phân l p kh c tính vi sinh c a th c khu n th i kháng vi khu n Edwardsilla ictaluri gây b nh cá tra t m u bùn .30 3.1.1 K t qu phân l p .30 3.1.2 K t qu t o stock th c khu n th phân l 3.1.3 K t qu ho t tính xâm nhi m .34 3.1.4 K t qu c 31 nh ph xâm nhi m c a th c khu n th vi khu n khác 36 3.2 Thí nghi m genome c a th c khu n th .38 3.2.1 Ki m tra x lý d li u thô 38 3.2.2 L p ráp h gen 45 3.2.3 Chú gi i c u trúc .47 HVTH: Nguy n Lê Hoài Nhân Trang vi Lu 3.2.4 Chú gi i ch 48 3.2.5 Phân tích phát sinh lồi 65 K T LU N VÀ KI N NGH 67 4.1 K t lu n 67 4.2 Ki n ngh 67 DANH M C CƠNG TRÌNH NGHIÊN C U 68 TÀI LI U THAM KH O 69 PH L C .i PH L C iii PH L C v PH L C vii PH L C ix PH L C xi PH L C xiii LÝ L CH TRÍCH NGANG xv HVTH: Nguy n Lê Hoài Nhân Trang vii Lu PH L C NG PH N M M PROKKA PVN03 B NG K T QU ORF 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 Nucleotide Nucleotide 12 323 577 2284 2789 3276 3967 4326 4751 4987 5765 5938 6330 6514 6694 7021 8322 9258 9725 9990 10156 10457 10692 11132 12544 15551 16158 16533 16700 17098 17358 17772 19665 21089 22314 22955 24097 24459 25135 326 565 2274 2652 3229 3974 4311 4739 4891 5763 5938 6120 6509 6660 6954 8325 9017 9707 9979 10175 10383 10702 10877 11329 15078 16090 16526 16703 17086 17331 17762 19655 21092 22312 22958 24100 24447 25133 26328 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân hypothetical protein hypothetical protein DNA primase/helicase hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang i Lu 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 26321 27397 31273 31721 32186 33604 34121 34593 34973 35524 36622 37143 38221 39556 39749 42055 42194 44068 44658 45572 46078 46271 46821 47793 48710 27397 31086 31701 32176 33604 34131 34585 34964 35464 36558 37131 38243 39546 39738 41776 42216 44041 44658 45563 46078 46314 46816 47777 48575 50698 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang ii Lu PH L C NG PH N M M PROKKA PVN04 B NG K T QU ORF Nucleotide Nucleotide 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS 687 1034 2176 2822 4042 5479 7372 7803 8048 8431 8608 9044 10000 13748 13927 14228 14403 14704 15106 15378 16068 16760 17606 17900 18051 18440 18622 18797 19668 19820 20248 20585 675 1037 2179 2820 4045 5469 7362 7776 8036 8434 8601 8976 9583 12534 13924 14106 14413 14630 14931 15360 15827 16763 17539 17866 18046 18230 18622 18795 19573 19808 20233 20592 21283 34 CDS 21330 21770 35 36 37 38 39 40 CDS CDS CDS CDS CDS CDS 21907 22285 23994 24233 24547 26709 22275 23982 24236 24547 26574 27491 HVTH: Nguy n Lê Hoài Nhân hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Ribonucleoside-diphosphate reductase large subunit hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang iii Lu 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 27507 28471 28973 29209 29724 30640 31241 33371 34558 34998 37036 37228 38531 39643 40216 41310 41810 42189 42643 43170 44598 45073 45688 49377 50446 28466 29016 29209 29715 30629 31224 33361 34474 34719 37025 37218 38553 39631 40152 41250 41801 42181 42653 43170 44588 45053 45501 49377 50453 51720 hypothetical protein hypothetical protein hypothetical protein hypothetical protein DNA primase/helicase hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang iv Lu PH L C NG PH N M M PROKKA PVN05 B NG K T QU ORF Nucleotide Nucleotide 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS 39 2267 2446 2747 2922 3223 3625 3897 4587 5279 6650 6944 7095 7484 7666 7841 8713 8865 9293 9630 10375 10952 11330 13039 13278 13592 15754 16552 17513 18015 18251 18766 19671 1052 2443 2625 2932 3149 3450 3879 4346 5282 6583 6910 7090 7274 7666 7839 8617 8853 9278 9637 10328 10815 11320 13027 13281 13592 15619 16536 17508 18058 18251 18757 19671 20261 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein DNA primase/helicase hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein 34 CDS 20288 22135 Anaerobic ribonucleosidetriphosphate reductase 35 36 37 38 CDS CDS CDS CDS 22113 22553 24591 24783 22274 24580 24773 26108 hypothetical protein hypothetical protein hypothetical protein hypothetical protein HVTH: Nguy n Lê Hoài Nhân Trang v Lu 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 26086 27198 27771 28865 29365 29744 30198 30725 32153 32628 33243 36932 38001 39277 39963 40310 41452 42098 43318 44755 46648 47150 47417 48106 48450 48833 49010 49446 50897 27186 27707 28805 29356 29736 30208 30725 32143 32608 33056 36932 38008 39275 39951 40313 41455 42096 43321 44745 46638 47052 47407 48097 48438 48836 49003 49378 49985 51670 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang vi Lu PH L C NG PH N M M PROKKA PVN06 B NG K T QU ORF 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 Nucleotide Nucleotide CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 556 893 1054 2712 3060 3619 3922 6521 6721 6936 7616 7777 8649 8966 9211 10341 10747 12132 12555 13959 14555 15500 15724 16086 16310 16597 16782 17203 18420 18922 20661 21068 21331 21774 22160 22891 23283 24002 24597 24946 25529 762 1060 2712 3056 3560 3903 6270 6730 6939 7619 7777 8649 8969 9247 9855 10760 12135 12347 13928 14489 15364 15727 16089 16313 16594 16782 17156 18420 18917 19986 20930 21229 21570 22121 22825 23046 23705 24595 24956 25464 26671 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang vii Lu 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 26685 27089 27639 28295 28592 28719 29069 29281 29478 29801 31087 32613 33335 33678 34886 35532 36146 36631 36939 37529 37657 38100 38386 27089 27490 28298 28591 28732 29072 29287 29481 29741 31090 32478 33338 33706 34889 35530 36146 36496 36942 37472 37654 38088 38369 38913 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang viii Lu PH L C NG PH N M M PROKKA PVN07 B NG K T QU ORF 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 Nucleotide Nucleotide CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 341 1499 2332 3893 4438 5073 5617 5898 6332 6514 7044 7490 7840 8456 9097 10280 10648 11508 12896 14245 14505 14699 14914 15254 15400 15844 16137 16945 17346 17764 18971 19479 19840 20730 21389 21610 22314 22865 23206 23505 1492 2284 3861 4441 5034 5600 5886 6329 6457 7047 7355 7840 8454 9100 10308 10651 11373 12899 14185 14508 14705 14917 15267 15397 15843 16140 16796 17346 17750 18906 19489 19838 20433 21152 21544 22275 22661 23104 23367 23774 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang ix Lu 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 24110 24490 25559 26056 27320 27694 27882 28163 28427 29302 29863 31444 31656 33031 33936 34544 34822 35142 36014 36172 36852 37061 37521 39888 40231 40735 41079 42731 43029 24424 25554 26056 27273 27694 27879 28166 28291 29236 29832 31236 31659 33044 33450 34580 34825 35142 36014 36175 36855 37070 37270 39869 40172 40731 41079 42737 42898 43235 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang x Lu PH L C NG PH N M M PROKKA PVN08 B NG K T QU ORF 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân Nucleotide Nucleotide 312 1470 2312 4468 5012 5293 5727 5909 6439 6885 7235 7851 8492 9675 10043 10903 12291 13640 13900 14094 14309 14649 15001 15142 15435 16243 16644 17062 18269 18777 19138 20028 20861 21571 22052 22441 22812 23409 23789 24858 1463 2264 4429 4995 5281 5724 5852 6442 6750 7235 7849 8495 9703 10046 10768 12294 13580 13903 14100 14312 14659 15014 15141 15438 16094 16644 17048 18204 18787 19136 19731 20450 21532 22050 22444 22674 23081 23723 24853 25355 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang xi Lu 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS CDS HVTH: Nguy n Lê Hoài Nhân 25355 26619 26993 27181 27462 27726 28601 29162 30743 30955 32330 33198 33749 34027 34347 35219 35377 36057 36266 36726 39093 39436 39940 40284 41936 42233 26572 26993 27178 27465 27590 28535 29131 30535 30958 32343 32749 33785 34030 34347 35219 35380 36060 36275 36475 39074 39377 39936 40284 41942 42103 42439 hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein hypothetical protein Trang xii Lu PH L C Hóa ch ng ng Trytic Soy Broth (TSB) c a hãng Himediang TSB 0.4% Agar (1000ml) Hóa ch t Kh ng Th tích N cu i TSB 30g 3% Agar 4g 0.4% ng Luria-Bertani (LB) 1.5% Agar (1000ml) ch nh pH =7 Hóa ch t Kh ng Th tích N cu i Pepton 10g 1% Cao n m men 5g 0.5% NaCl 5g 0.5% Agar 15g 1.5% SM buffer (50ml) Hóa ch t Kh ng Th tích N cu i NaCl 0.29 g 100 mM MgSO4.7H2O 0.1 g mM Tris.HCl 1M (pH = 7.5) 2.5 mL 50 mM Gelatin 2% 0.25 mL 0.002% ng d tr th c khu n th (50 mL) Hóa ch t Th tích SM buffer Glycerol Thi t b d ng c 2.1 N 40 mL 10 mL cu i 80% 20% Thi t b STT Tên nt Ký hi u Cân Máy l c n nhi t Jisico N i h p ti t trùng HIRAYAMA HV85 T c y vơ trùng Huy Hồng T l nh SANYO HVTH: Nguy n Lê Hoài Nhân Sartorius Trang xiii Lu N i h p ti t trùng HIRAYAMA HV85 T MEMMERT T âm sâu -700C Sanyo Microway SHARP 800WATT m 10 Máy ly tâm l nh Centrifuge 5415R c a Eppendoft 11 B p khu y t gia nhi t Stuart 12 Lị vi sóng 13 B n nhi t 14 Máy l c vòng 15 Máy vortex 16 T s y 2.2 MULTI MB45 D ng c STT Danh m c ng nghi m lo i C c th y tinh Bình tam giác ng nghi m D ng c hút m u tuýp HVTH: Nguy n Lê Hoài Nhân STT Danh m c ng Falcon 15ml 50ml petri phi 60 90 n Eppendoft 1.5ml 10 Màng l c 0.22um Trang xiv Lu LÝ L CH TRÍCH NGANG H tên: Nguy n Lê Hoài Nhân nh a ch liên l c: 20/130/2A Tân Th Th i gian , Qu n Tân Phú, TP.HCM o 2012 - 2016 2017 - 2021 i h c Công Nghi p Th c Ph m TPHCM i h c Bách Khoa TPHCM o Hình th c ng o Cơng ngh sinh h c Chính quy K Cơng ngh sinh h c Chính quy Th Q TRÌNH CƠNG TÁC Th i gian 2016 Ch c v 2020 Cơng Ty TNHH Hóa Vi t Kinh doanh n Công Ty TNHH Khoa h c k thu t H Kinh doanh HVTH: Nguy n Lê Hoài Nhân Trang xv ... TÀI Phân l p phân tích genome th c khu n th kháng vi khu n gây b nh quan tr ng cá tra II NHI M V VÀ N I DUNG Phân l p kh c tính vi sinh c a th c khu n th Edwardsiella ictaluri gây b nh cá tra. .. n th Phân tích genome th c khu n th kháng Aeromonas hydrophila phân l p Vi l p Vi i genome c a PVN02 phân 2020 Phân tích genome th c khu n th kháng Edwardsiella ictaluri phân l p Vi t Nam hi... with the bacteriophages previously present on NCBI HVTH: Nguy n Lê Hoài Nhân Trang iii Lu L Phân l p phân tích genome th c khu n th kháng vi khu n gây b nh quan tr ng cá tra v is u c a ng d n c