Luận văn : NGHIÊN CỨU HIỆN TRẠNG NHIỄM BỆNH TSWV (Tomato spotted wilt virus) TRÊN CÂY ỚT BẰNG KỸ THUẬT ELISA VÀ BƢỚC ĐẦU XÂY DỰNG PHƯƠNG PHÁP CHẨN ĐOÁN BẰNG KỸ THUẬT RT - PCR part 8 pot

59

|P12 |TGCTTGAATTTGTCATGTCCAAGG | 53 | 59 | -49 | 42 | 26-49 |
|M4 |CCTGCAATAGAGAGGAATAATCGC | | 56 | -50 | 46 | 923-946 |
| | | | | | | |
|P14 |GCAGATGCTTGAATTTGTCATGTCC | 53 | 59 | -50 | 44 | 21-45 |
|M6 |GCAATAGAGAGGAATAATCGCTCC | | 55 | -49 | 46 | 920-943 |
| | | | | | | |
|P13 |ATGCTTGAATTTGTCATGTCCAAGG | 53 | 59 | -50 | 40 | 25-49 |
|M6 |GCAATAGAGAGGAATAATCGCTCC | | 55 | -49 | 46 | 920-943 |
| | | | | | | |
|P12 |TGCTTGAATTTGTCATGTCCAAGG | 53 | 59 | -49 | 42 | 26-49 |
|M6 |GCAATAGAGAGGAATAATCGCTCC | | 55 | -49 | 46 | 920-943 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M9 |CCTGCAATAGAGAGGAATAATCG | | 53 | -46 | 43 | 924-946 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M8 |CTGCAATAGAGAGGAATAATCGC | | 53 | -46 | 43 | 923-945 |
| | | | | | | |
|P36 |CATGTAGCGCAGAGTGATCC | 52 | 51 | -42 | 55 | 187-206 |
|M12 |TGCAATAGAGAGGAATAATCGC | | 52 | -45 | 41 | 923-944 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M12 |TGCAATAGAGAGGAATAATCGC | | 52 | -45 | 41 | 923-944 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M11 |CAATAGAGAGGAATAATCGCTCC | | 52 | -46 | 43 | 920-942 |
| | | | | | | |
|P28 |AGAGTGATCCATCGGAAGCC | 52 | 54 | -44 | 55 | 197-216 |
|M9 |CCTGCAATAGAGAGGAATAATCG | | 53 | -46 | 43 | 924-946 |

| | | | | | | |
|P28 |AGAGTGATCCATCGGAAGCC | 52 | 54 | -44 | 55 | 197-216 |
|M8 |CTGCAATAGAGAGGAATAATCGC | | 53 | -46 | 43 | 923-945 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M6 |GCAATAGAGAGGAATAATCGCTCC | | 55 | -49 | 46 | 920-943 |
| | | | | | | |
|P30 |TAGCGCAGAGTGATCCATCG | 52 | 54 | -44 | 55 | 191-210 |
|M12 |TGCAATAGAGAGGAATAATCGC | | 52 | -45 | 41 | 923-944 |
| | | | | | | |
|P28 |AGAGTGATCCATCGGAAGCC | 52 | 54 | -44 | 55 | 197-216 |
|M12 |TGCAATAGAGAGGAATAATCGC | | 52 | -45 | 41 | 923-944 |
| | | | | | | |
|P28 |AGAGTGATCCATCGGAAGCC | 52 | 54 | -44 | 55 | 197-216 |
|M11 |CAATAGAGAGGAATAATCGCTCC | | 52 | -46 | 43 | 920-942 |
| | | | | | | |
|P32 |CAGAGTGATCCATCGGAAGC | 52 | 53 | -43 | 55 | 196-215 |
|M5 |GCCTGCAATAGAGAGGAATAATCG | | 56 | -50 | 46 | 924-947

Từ kết quả trên chúng tôi đã chọn ra một cặp primer để chạy Nested PCR
 Thông số primer:
 Nhiệt độ Tm: 54
o
C và 55
o
C
 % GC: 55 % và 46%
 Nhiệt độ bắt cặp Ta: 53
o
C

 Vùng bắt cặp: 197-216 và 920-943
 Trình tự primer 5’ – 3’
P28 5’-AGAGTGATCCATCGGAAGCC-3’
M6 5’-GCAATAGAGAGGAATAATCGCTCC-3’

60

Phần V
Kết luận và đề nghị
5.1 Kết luận
Qua điều tra và chẩn đoán, chúng tôi thu nhận đƣợc các kết quả nhƣ sau:
 Tỷ lệ nhiễm TSWV theo địa bàn điều tra
- Xã Nhuận Đức: 12,1%
- Xã Phƣớc Thạnh: 12,5%
- Xã Lộc Hƣng: 0%
 Tỷ lệ nhiễm TSWV theo triệu chứng
- Lá khảm vàng xanh, cong: 37,5%
- Lá chấm vàng xanh, nhăn nheo, co lại, dày, giòn: 57%
- Lá khảm nặng có chấm đen, lủng lỗ:53%
- Khảm vàng nhạt trên cả rìa lá, lá nhỏ: 25%
 Tỷ lệ nhiễm trên trái
- Trái nhiễm: 33%
- Không nhiễm: 67%
Nhƣ vậy khi điều tra trên đồng ruộng thì chúng ta có thể đánh giá bệnh TSWV dựa
trên các triệu chứng trên. Đánh giá bệnh dựa vào giống và tuổi cây thì chƣa thực hiện
đƣợc vì bà con nông dân chƣa thực sự quan tâm đến giống mà họ đang trồng, và thời
gian nghiên cứu thì lại rơi vào cuối vụ, tất cả đều cho trái nhƣ nhau.
5.2 Đề nghị
 Tiếp tục nghiên cứu mức độ gây hại của TSWV theo giống và độ tuổi.
 Nghiên cứu khả năng lây lan của TSWV trên các đối tƣợng khác.

 Sử dụng cây chỉ thị để hỗ trợ cho việc chẩn đoán.
 Vì mức độ nhiễm của TSWV chƣa cao, lƣợng RNA virút ly trích đƣợc còn quá
thấp không thể phát hiện đƣợc bằng PCR thông thƣờng nên chúng tôi đề nghị
phƣơng pháp chẩn đoán TSWV bằng Nested PCR với cặp primer đang sử dụng
và cặp primer đƣợc vừa thiết kế. Hiệu quả của phƣơng pháp này sẽ đƣợc chứng
minh khi chúng tôi có điều kiện để thực hiện phản ứng Nested PCR với cả hai
cặp primer đã đƣợc đề cập ở phần trên.
 Xây dựng quy trình xác định TSWV bằng kỹ thuật Realtime – PCR.
 Thực hiện thu RNA bằng kỹ thuật siêu ly tâm theo khuynh độ đƣờng.
61

Phần VI
TÀI LIỆU THAM KHẢO

TIẾNG VIỆT
1. Nguyễn Ngọc Bích, 2004, Luận văn Thạc sĩ Khoa học Nông nghiệp: Bước đầu
nghiên cứu một số bệnh virus trên thuốc lá của tỉnh Tây Ninh.
2. Vũ Triệu Mân, 2003, Chẩn đoán nhanh bệnh hại thực vật. Nhà xuất bản Nông
nghiệp, Hà Nội, trang 46-50.
3. Vũ Triệu Mân – Lê Lƣơng Tề, 1999, Bệnh Vi khuẩn và Virus hại cây trồng. Nhà
xuất bản Giáo Dục, trang 143.
4. Lƣơng Nguyễn Thu Tâm,1996-2001, Luận văn tốt nghiệp Ngành Nông học: Tìm
hiểu ảnh hưởng của các loại phân bón lá đến khả năng sinh trưởng, phát triển và
năng suất cây ớt (Capsicumannum L.) trồng trong vụ mùa mưa tại phường Thống
Nhất-TP. Biên Hoà-Tỉnh Đồng Nai.
5. Phạm Đức Toàn, 1996-2000, Luận văn tốt nghiệp Ngành Nông học: Sử dụng kỹ
thuật ELISA để chẩn đoán bệnh Tristeza trên cây cam quýt tại quận 9, Thành Phố
Hồ Chí Minh.
6. Nguyễn Hữu Trúc,1995 -2000, Luận văn tốt nghiệp Ngành Nông học: Hiệu lực của
một số thuốc Bảo vệ Thực vật phòng trừ bệnh thán thư (Colleto trichum spp.) trên

ớt (Capsicumannum L.). Bước đầu xây dựng qui trình phòng trừ tổng hợp bệnh
thán thư ớt.
7. Nguyễn Văn Tuất, 2002, Kỹ thuật chẩn đoán và giám định bệnh hại cây trồng. Nhà
xuất bản Nông nghiệp, Hà nội, trang 69.
8. Bùi Cách Tuyến, 1998, Bản dịch tiếng việt, Tài liệu hướng dẫn đồng ruộng, Bệnh
hại cây ớt. Nhà xuất bản Nông nghiệp, trang 42 – 56.
9. Bùi Trang Việt, 2002, Sinh học phân tử, ĐH Quốc gia TP. HCM Khoa Sinh học -
Trƣờng ĐHKHTN.

TIẾNG NƢỚC NGOÀI
10. Alfred Nobel Dr. Hercules, CA 94547 USA, Inc, 2000, Aurum Total RNA Mini
Kit Instruction Manual Bio-Rad Laboratories, trang 26-27.
62

11. Biowave vol.6 No.23 2004, tomato spotted wilt virus.
<www.bric.postech.ac.kr/webzine/content/review/virology/2005/v7n12/pv0616.pdf >
12. Chemicon International, Inc - 28820 Single Oak Drive - Temecula, CA 92590,
Introduction to Antibodies - Enzyme-Linked Immunosorbent Assay (ELISA)
<
13. G.A. Dafalla, Plant Pathology Centre, Faculty of Agricultural Sciences,University
of Gezira, Wad Medani, Sudan.
<www.iita.org/info/virology/pdf_files/18-24.pdf>
14. Introduction to Microbiology: Virus Genomes, 2005.
<
15. J.Morris, Central Science Laboratory, Sand Hutton, York, Y041 1LZ, UK.
<www.csl.gov.uk/science/organ/ph/diagpro/tospovirus.pdf>
16. John R. Crowther, 2001, The Elisa Guidebook – Volume 149, trang 12 – 34.
17. Ken Pernezny-Everglades research and Education Center University of Florida,
Belle Glade; Pamela D. Roberts-Southwest Florida Research and Education Center
University of Florida, Immokalee; John F. Murphy-Auburn University, Auburn,

Alabama; Natalie P. Goldberg-New Mexico State University, Las Cruces;
Compendium of pepper Diseases; 2003; The American Phytopathological Society.
18. Lindsey Irons and Emily Sims, tomato spotted wilt virus.
<www.rhodes.edu/biology/glindquester/ viruses/pagespass/tswv/tswv.html>
19. Marcelo Eiras1, Renato O. Resende2, Alexandre A. Missiaggia2 & Antônio c. de
Ávila3, Centro de Sanidade Vegetal, Instituto Biológico, CEP 04014-002, São
Paulo, SP, Departamento de Biologia Celular, Universidade de Brasília, CEP
70910970,Brasília, DF; 3Centro Nacional de Pesquisas de Hortaliças -
EMBRAPA, Cx. Postal 218, CEP 70359-970, Brasília, DF, RT – PCR and dot blot
hybridization methods for a universal detection of tospoviruses, 2001.
<www.scielo.br/pdf/fb/v26n2/a09v26n2.pdf>
20. Michael Blaber, 1998, BCH5425 Molecular Biology and Biotechnology
<www.wine1.sb.fsu.edu/bch5425/lect30/lect30.htm - 14k>
21. Peter Schreier, Prof. Dr. Martin Hülskamp, 2002, Molekulare Charakterisierung
des Nukleokapsidaufbausvon TSWV (Tomato Spotted Wilt Virus)
<www.ub.uni-koeln.de/neuerwerbung/00/17/2003_01_14.html - 9k>
63

22. P. Sreerama Kumar, ProjectDirectorate of Biological Control (ICAR), P. B. No.
2491, H. A. FarmPost, Hebbal, Bellary Road, Bangalore 560 024, Karnataka, India.
PlantDis. 84:1151, 2000; published on-line as D-2000-0816-03N, 2000.Accepted
for publication 30 June 2000.
<www.cnr.berkeley.edu/garbelotto/ downloads/davidson2002.pdf>
23. Ray Cerkauskas, Visiting Scientist from Agriculture and Agri-Food Canada.
Edited by Tom Kalb. Photos by L.L. Black and E.L.Shannon. Published by
AVRDC – The World Vegetable Center; P.O. Box 42, Shanhua; Taiwan 741;
ROC. 2004.
<www.avrdc.org/pdf/pepper/TSWV.pdf>
24. Sambrook và Russell, Third edition, Molecular Cloning – volume 2, trang 8.47.
25. Theo SK&ĐS, 2005, Vị thuốc từ ớt.

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26. Vernon E Coyne, M Diane James, Sharon J Reid and Edward P Rybicki,Molecular
Biology Techniques Manual, Third Edition, 1996.
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27. <www.apsnet.org/pd/pdfs/2002/0926-01R.pdf>.
28. <www.promega.com/guides/rna_guide/amplifyingrna.pdf>.













PHẦN VII
PHỤ LỤC

Phụ lục 1: Trình tự Nucleotide của genome TSWV
1 agagcaattg tgtcagaatt ttgttcataa tcaaacctca cttagaaaat cacaatactg
61 taataagaac acagtaccaa taaccataat gtcttcaagt gtttatgagt cgatcattca
121 gacaagagct tcagtctggg gatcaactgc atctggtaaa gctgttgtag attcttactg
181 gattcatgaa cttggtactg gttctcaact agttcagacc cagctgtatt ctgattcaag
241 aagcaaagta gtcctttggc tatactgcaa agtagggatc ttccctgtga agaagaagag
301 atttctttct cagcatgtgt atatccctat ttttgatgat attgatttta gcatcaatat
361 tgataactct gttctggcac tatctgtttg ctcaaataca gtcaatgcta acggagtgaa

421 acatcaaggt catttgaagg ttttgtctcc tgcccagctc cactctattg aatctatcat
481 gaacagatct gatattacag accgattcca gctccaagaa aaagacataa ttcccaatga
541 caaatacatt gaagctgcaa acaaaggctc tttgtcttgt gtcaaagagc atacctataa
601 gatcgagatg tgctataatc aggctttagg caaagtgaat gttctatctc ctaacagaaa
661 tgtccatgaa tggctgtaca gtttcaagcc aaatttcaat caagttgaaa gcaacaacag
721 aactgtaaat tctcttgcag tgaaatctct gctcatgtca gcagaaaaca acatcatgcc
781 taactctcaa gcttccactg attctcattt caagctgagc ctctggctaa gggttccaaa
841 ggttttgaag caggtttcca ttcagaaatt gttcaaggtt gcaggagatg aaacaaacaa
901 aacattttat ttatctattg cctgcattcc aaaccataac agtgttgaga cagctttaaa
961 cattactgtt atttgcaagc atcagctccc aattcgcaaa tgcaaagctc cttttgaatt
1021 atcaatgatg ttttctgatt taaaggagcc ttacaacatt gttcatgacc cttcataccc
1081 caaaggatcg gttccaatgc tctggctcga aactcacaca tctttgcaca agttctttgc
1141 aactaacttg caagaagatg taatcatcta cactttgaac aaccttgagc taactcctgg
1201 aaagttagat ttaggtgaaa gaaccttgaa ttacagtgaa gatgcctaca aaaggaaata
1261 tttcctttca aaaacacttg aatgtcttcc atctaacaca caaactatgt cttacttaga
1321 cagcatccaa atcccttcat ggaagataga ctttgccaga ggagaaatta aaatttctcc
1381 acaatctatt tcagttgcaa aatctttgtt aaagcttgat ttaagcggga tcaaaaagaa
1441 agaatctaag gttaaggaag cgtatgcttc aggatcaaaa taatcttgct ttgtccagct
1501 ttttctaatt atgttatgtt tattttcttt ctttacttat aattatttct ctgtttgtca
1561 tctctttcaa attcctcctg tctagtagaa accataaaaa caaaaaataa aaatgaaaat
1621 aaaattaaaa taaaataaaa tcaaaaaatg aaataaaaac aacaaaaaat taaaaaacga
1681 aaaaccaaaa agacccgaaa gggaccaatt tggccaaatt tgggttttgt ttttgttttt
1741 tgttttttgt tttttatttt ttattttatt tttattttat tttattttta ttttattttt
1801 attttattta ttttttgttt tcgttgtttt tgttatttta ttatttatta agcacaacac
1861 acagaaagca aactttaatt aaacacactt atttaaaatt taacacacta agcaagcaca
1921 agcaataaag ataaagaaag ctttatatat ttataggctt ttttataatt taacttacag
1981 ctgctttcaa gcaagttctg cgagttttgc ctgcttttta accccgaaca tttcatagaa
2041 cttgttaaga gtttcactgt aatgttccat agcaacactc cctttagcat taggattgct
2101 ggagctaagt atagcagcat actctttccc cttcttcacc tgatcttcat tcatttcaaa
2161 tgctttgctt ttcagcacag tgcaaacttt tcctaaggct tccttggtgt catacttctt

2221 tgggtcgatc ccgaggtcct tgtattttgc atcctgatat atagccaaga caacactgat
2281 catctcaaag ctatcaactg aagcaataag aggtaagcta cctcccagca ttatggcaag
2341 tctcacagac tttgcatcat cgagaggtaa tccataggct tgaatcaaag gatgggaagc
2401 aatcttagat ttgatagtat tgagattctc agaattccca gtttcttcaa caagcctgac
2461 cctgatcaag ctatcaagcc ttctgaaggt catgtcagtg cctccaatcc tgtctgaagt
2521 tttctttatg gtaattttac caaaagtaaa atcgctttgc ttaataacct tcattatgct
2581 ctgacgattc tttaggaatg tcagacatga aataacgctc atcttcttga tctggtcgat
2641 gttttccaga caaaaagtct tgaagttgaa tgctaccaga ttctgatctt cctcaaactc
2701 aaggtctttg ccttgtgtca acaaagcaac aatgctttcc ttagtgagct taaccttaga
2761 catgatgatc gtaaaagttg ttatatgctt tgaccgtatg taactcaagg tgcgaaagtg
2821 caactctgta tcccgcagtc gtttcttagg ttcttaatgt gatgatttgt aagactgagt
2881 gttaaggtat gaacacaaaa ttgacacgat tgctct
Hình pl.1a: Trình tự Nucleotide genome S (Short)




1 agagcaatca gtgcatcaga aatataccta ttatacattt tgctaagaat caatcaacta
61 cattacacaa gctcctctac cttaggctgt tgaactcgaa atgttgactc ttttcggtaa
121 caagaggcct tctaagtctg taatggcagt gttgaagtct caaaaccatg gtcttcttct
181 gatgaaaagc ttgctttaac ccggaaagga tgaaggtcct ttagtttcac ttgctaaaca
241 caaagccatg gatgcatcca aaggaaagat actgttaaac attgagggaa catcttcctt
301 tggaacctat gaatctgatt ccatcacaga gtcagaaggt tatgatcttt ctgctagaat
361 gatagtagat acaaaccatc atatctcaaa ctggaaaaat gatctttttg ttggcaacgg
421 aaagcaaaat gctaataagg ttatcaagat ctgtccgact tgggacagca gaaaacaata
481 catgatgatt tccagaattg tgatatgggt ctgccccact ataccaaacc ctacaggaaa
541 acttgtggtt gctttaattg atcccaacat gccatctgga aagcaagtca tcctgaaggg
601 tcaggggaca ataactgatc ctatctgctt tgttttttat ctgaactggt ctattccgaa
661 gatgaacaac accccagaaa actgttgtca gctgcatttg atgtgtagcc aagaatacaa
721 gaaaggggtt tcttttggta gtgtcatgta ttcttggaca aaagagtttg gtgattcacc

781 cagagctgat aaagacaaaa gttgtatggt tatacctcta aacagggcca ttagagctag
841 atctcaggca ttcattgaag cctgcaagct gataattcct aaaggaaaca gtgagaagca
901 gataaaaaaa cagcttaaag aactgagctc aaatcttgag agatcagttg aagaggaaga
961 ggaaggaatt tctgacagtg ttgctcagtt atcctttgat gaaatatagt tctttaaata
1021 tcacttattt aagcttaaat ttctgtctat tttgcatttt gaatccaaaa acccaaacaa
1081 aaaacgaaaa caaaaaagag aaaaaaacaa aaaatcaaac caaaaaacaa aaacaaaata
1141 aggctgaaaa gcaaactttg gtccgaagac tttttttgtt gttttttgtt tatttttatt
1201 tttttgtttg ttttttgttt attttatatt tcgtttttat tagttaatga ttgattttaa
1261 agatttttct atatatataa tcctgctaat atagaagatt gaatcaaatt taatctgtga
1321 caagcatctt cagacaaggt gagagaaatc cataggtggc cttcttcttg tcattgtatc
1381 tttcattaac ataggggctt tgatctcagg ttcatcatca tcctctatct tggatctaga
1441 tttataagat tcattcttta catatccttt acaaatggat gtcagaatag aacagaaata
1501 agtcacaaga aaaatgaatg caataagcag taccactctg atagtatcaa aaaatgagcc
1561 aaagtaactt gcaatgaaat tgaatggact cttaatataa tcccagaagc cccatgctga
1621 agaatcagaa ttatattgtt gttcttcatg agcatactca tcattttgat ctattatatt
1681 ctctggttct tctacaataa cattattaac caaaacttcc acagagatat ccggattgcc
1741 ttctggatac aatgtcattt tcttcttgtc cggattggct gaacaaaaca ttgttatatt
1801 gtatttatta gatccttttt taacagctag ctgataagta gataaagagc aagcatctat
1861 agaaattgca gtagaaaatg tcaaatctga gaaaaattct aaaaggcaag ataaaccttg
1921 gccgcataga agacagccat tgcaatttaa gcttgttgaa gttatggaag gtttcttagg
1981 agcaacttta aaaagatcag atggaagatc aactaccatt ttcagtttcc ctagactaaa
2041 agattttgcc agaaaaaaac tagaaaaatc tttgaaacta acaggaatat ctgatatttg
2101 ctctaaacca gatctaaacc tgtatgtgtc gtatccacat gttttgatag tgactgattt
2161 tttccctatt gctgcacaat cccaagacat gtcatctcct tctagagttt tcttagtaaa
2221 aataggcact ccatcatggg tcaattgtgg atgaccaaac attttcacag gatcattcaa
2281 gtttgcaata tttccagagt aaatatgact gtcgggtcca tgagctatta gttcacctat
2341 agtgatacca tcattatgca aatctgcttg tatatcagct tgaaacaatg tattttcata
2401 aggaacctct tcagtaatcc ttgagcattg acctcccaaa ataccggaaa tacaaacatc
2461 tgctactata gttgacttaa gcactgaata aattctgtat gatttgtcca tatcataaat
2521 atttcgacag aatccgcatg tagcaccctc attaattgca aaacaccaag cttcttcaca

2581 tccccaataa gaagttggtg ttacacaaaa atcttggaaa cctgttaaag cttgattttt
2641 cctgcaagtg tcgcagtttc ctgtacaagt ggaataaaaa tccgtgtggg tgctttggat
2701 gggagctgtc gtatattttt ctgacacttc ataatgaatt cccacacttt tgatataaat
2761 cacaaatttt ttggctgttt ctgaggtctt gtcatttagc atgaatatag ttcctcctcc
2821 tcctaaaaga gattgttcta tcatatatct atatttcccg tctacaacag aatcaaagat
2881 taatgattgc ctgggcagaa tgttctctgg tatgcttgtt tcgtttttgt ttaaggagtc
2941 tccggcaacc attccagaaa ttttgccctg acggttatag gaatctatta taccatttct
3001 caatctctta gcctcataca gaccaactga tgttaaccct aaagagcttc ctgtataatc
3061 cgaaaaccca ttgtacaaag gtttgtttcg gaataaatct aggtattcgc aacctaatct
3121 gcattttata agattatcaa tagatgggcg tgcaatgcaa ttctggttct ctatgcaatc
3181 attaggacct tctataacaa tattagtgcc aaagatactc tctatgatct tatcttcttt
3241 tacattgcag taacattgat ctttttcaga gcatttttca aatttgcttg taaccaaaaa
3301 tggacagcct ggaacataaa agcatccact caaacattgg gttgtttgag ccatagacat
3361 gggcatctga gacaaaatga ctaaacctat caaaatttcg gtcacaaatt ttagcaaact
3421 caagctcagc ttagtgttca ctattagatg gaaccattcc atgctagtcc acttatgttt
3481 gttgtagtca tgatctgctt ctttggatag tatgggacac tctgaagaat gctcttttga
3541 agctttgctt ttgttgcaaa tgcagacttt agtacactca tgtgtgacta tgcacagatt
3601 gccgcagtta gaacatttta atgggaaata tttccataag caatttatga gcaataagac


3661 agggtatgta atcaagccca taagatcata ccagagaaag agaggtttag tcgtcttgtt
3721 cactaaccat cggataggga aatagatcaa caaagctatc aatatcaacc ttatccaaga
3781 aaaattgatg caggctgttt gcttataaat actttttgaa tatttgatta tgcaatctct
3841 gactcttttg tttgtttttg gtattttggc tgatttgtca ccacacaaga gattgtgttc
3901 accatccaac atttcctcag taaaagtgat acttgctgat ccagaaaaag atataacctt
3961 gtgttccaca ttttcaccag gtttttttat caaataaccc atgatcttct cagggctagt
4021 gatgcttaca gtatagggat ttgcgaagtt tgatttagtt attttgcagt caccggataa
4081 ctttacagtt tgtaatgata ctgttccatt agtggggtat gagttgtaag ttataggata
4141 attatcttgt gtcaggcttt ctgaaatgaa gaattttcct cctactgaaa agtgtctttt
4201 cttgtctagc ttggtaatgg gaataaccgg gactttggag aatctctttg gcaaatttaa

4261 agaattatca catttttcta aaccttctgc tgaatcagaa acacaggaat atatgacacc
4321 attgttttca acttgataat aaacattata agtggatatc ccttttatct cacattttaa
4381 tgaagaagca ttcaagcaat tgttgggaag atccaaaaca gagattgttt tttgcgttgt
4441 tggctcagca gaaataggga tggttgattt ttcttctcgt atctgacggg ttccaggagt
4501 ccgagattct agcatcagat tagttaaagt ctctaagata gcttcgcgtt gaatcgatgc
4561 agcagtgggt acctcattct cagcagaatc atcataaatc tcaggatgat ctccacgaat
4621 tatttctact ttagcatctg tggctctgaa gatcaagaat gccaacaaaa cagaactcag
4681 ggcaattgtg aaaagactca cttttaccac tagttctagt agttttagaa ttctcatctt
4741 agatgtctac ccagattaca atggttgtgt gattaatttc aagatgtctg gattaaggtt
4801 tttgtttgca ctgattgctc t

Hình pl.1b: Trình tự Nucleotide genome M (Medium)

1 agagcaatca ggtaacaacg attttaagca aacatgaaca tccagaaaat acaaaaatta
61 atagaaaatg gaaccacttt actgttgtct attgaggatt gtgtaggttc taaccacgat
121 ctagctttgg atttacataa gagaaatagt gatgagatcc cagaagatgt gattataaat
181 aataatgcaa aaaattatga gacaatgaga gagttaattg tcaaaatcac tgctgatggt
241 gaaggactaa acaaagggat ggcaactgtg gatgtcaaaa agctaagtga gatggtctct
301 ctgtttgagc aaaaatacct agaaacagag ttagcaaggc atgacatttt tggagagctg
361 atctccaggc acctgagaat aaagcccaaa caaagaaatg aagtggagat agagcatgca
421 ctaagagaat atctggatga actcaacaaa aagtcctgca ttaacaagct ctctgatgat
481 gagtttgaga gaataaataa agaatatgta gcaactaatg ccacccctga taactatgtg
541 atatataaag aatcaaaaaa cagtgagctt tgtttaatca tttatgattg gaaaatatct
601 gtcgatgcca ggactgaaac caaacaatgg agaaatacct acaagaatat ttggaaatct
661 ttcaaagata taaaagtgaa tggaaagcca ttcctggaag agcatcctgt tttcgtttct
721 atagttatat tgaaacctat tgctgggatg ccaatcactg ttactagtag cagggttttg
781 gagaaattcg aagattctcc atcagcattg cacggagaaa gaataaagca tgctaaaaat
841 gccaaattgc taaatatttc ttatgttggg caaatagttg gaaccacacc cacagtggtg
901 agaaactatt atgcaaacac tcaaagaatc aaatctgaag tcagaggaat cttaggtgat
961 gattttggat ctaaagatgt gtttttcagt cactggacca gcaaatacaa agaaagaaat

1021 cctactgaga tagcctattc cgaagatatt gaaagaataa ttgattcact tgttacagat
1081 gaaatcccta gagaggaaat aatacatttt ttgtttggaa atttctgttt ccacattgaa
1141 acaatgaatg accagcatat agctgacaaa tttaaagggt accaaaactc ttgtatcaat
1201 ttaaaaatag agccaaaagc tgatttagct gatttgaaag accacttaat ccaaaagcag
1261 caaatatggg aatctctgta tggaaaacac cttgagaaga tcatgcttag aattagagaa
1321 aaaaagagaa aagaaaaaga gatacctgac ataaccacag cttttaacca gaatgctgct
1381 gaatatgaag aaaggtatcc taactgtttc aatgatctct ctgaactaaa actaactttc
1441 catgacttgg tccccagttt gaagatagaa ttgagctcag aggtagatta caacaacgca
1501 attattaaca agtttcggga gagcttcaaa agttcttcaa gggttattta taatagccca
1561 tatagtagca taaataacca aacaaataaa gcaagagata taacaaactt agttagactg
1621 tgtttagcag agctaagttg tgatacaacg aaaatggaaa agcaggaact tgaagatgaa
1681 atagatataa acaccgggag tatcaaagtt gagagaacaa aaaagtctaa agaatggaat
1741 aagcaaggtt cgtgtttaac cagaaacaaa aatgaatttt gcatgaaaga tacaggcagg
1801 gagaacaaaa ctacctattt taaaggctta gcagtaatga atataggaat gagttctaag
1861 aaaagaattc taaaaaaaga agaaataaaa gaaaggatct ctaaaggctt ggaatatgat
1921 acctctgaaa ggcaggctga cccaaatgat gattactcaa gtatagacat gtcttctctg
1981 actcatatga aaaaactgat aaggcatgac aatgatgata gcttaagtgg taaaagattt
2041 aagggctctt tttttctact tcataatttt aatataatag aggatggtaa gatcacatct
2101 gttttcaata attatgctaa aaatcctgaa tgcttgtaca ttcaagattc agtactgaag
2161 actgaattag agacttgcaa aaagataaac aaattatgca atgatctagc catttaccat
2221 tactctgaag acatgatgca attctccaaa ggtttaatgg tggctgacag gtacatgact


2281 aaagaaagtt tcaagatatt aaccacagca aatactagca tgatgctatt agcattcaaa
2341 ggggatggaa tgaacaccgg aggatcggga gttccttaca tagcattgca tatagtggat
2401 gaagacatgt cagatcattt taacatatgt tatactaaag aaatttatag ctatttccga
2461 agtggtagta attacattta tataatgagg ccgcagagac taaaccaggt gaggctgctg
2521 aggcttttca aaacgcctag taaagttcct gtatgttttc cacaattttc aaagaaagct
2581 aatgaaatcg gaaaatcgct gaaaaataaa gatatagaaa aagtaaatct cttttctatg
2641 acaatgactg taaaacagat attaataaat attgtgtttt catctgtcat gataggaact

2701 gtgacaaagc tcagtagaat gggaattttt gatttcatgc ggtatgcagg ttttttgcga
2761 ctatccgatt attctaacat aaaagaatac attagagaca aatctgatcc tgatataact
2821 aactgtggca gatatctatt tcgtaatgga atcaaaaaac tattgttcag aatggaagat
2881 ctcaatttaa gcacaaatgc caagcctgtt gttgtggacc acgaaaatga tattatagga
2941 gggataacaa acttgaatat aaaatgtcct ataacaggat caactctact gacacttgag
3001 gacctgtaca ataatgttta tttggctatt tatatgatgc ctaaatcact gcacaatcat
3061 gttcacaatc taacaagctt attaaatctc cctgctgagt gggagctaaa gttcagaaaa
3121 gaattaggtt tcaacatatt tgaagacata taccctaaga aagcaatgtt tgatgacaaa
3181 gacctattct ccataaatgg agctttgaac gtgaaagcat tatctgatta ctatctagga
3241 aatatagaaa atgttggttt aatgagatca gaaatagaaa ataaagaaga tttcctaagc
3301 ccttgttata aaatatctac tttaaaatct tcaaaaaaat gctcacaatc aaacattata
3361 agtactgatg agataataga gtgtcttcag gatgcaaaga ttcaagatat agaaaattgg
3421 aaaggaaata acctagctat tataaaaggg cttataagaa cctacaatga ggagaaaaat
3481 agattggtgg aattttttga agataattgt gtcaattcat tatatcttat agaaaagctt
3541 aaagagataa ttagtagtgg atcaataact gtagggaaat ctgtaacatc taaattcata
3601 agaaacaatc atcctttaac agtagaaaca tatctcaaaa caaaactata ttatagaaat
3661 aatgtaacag ttttaaagtc taaaaaagtg tcagaggagc tctatgacct tgtaaaacag
3721 ttccatgaca tgatggaaat agacctagat tctgttatga acctcgggaa aggtacagaa
3781 ggaaaaaaac tcacattctt gcagatgctt gaatttgtca tgtccaaggc taaaaatgtc
3841 accgggtctg tagattttct agtttctgtt tttgaaaaaa tgcagagaac caaaacagac
3901 agagaaatat acttgatgag catgaaagtg aaaatgatgc tttattttat agagcataca
3961 ttcaaacatg tagcgcagag tgatccatcg gaagccatat ctataagtgg agacaataaa
4021 ataagagcac tttctacgtt atctttggac acaatcacgt cttacaatga tattttaaac
4081 aaaaattcaa agaagtcaag attggctttc ctatctgcag atcagtcgaa atggtcggca
4141 tcaggcctta ccacctataa atatgtttta gctatcatat taaatccaat tttaactact
4201 ggtgaagcta gcttaatgat agaatgcatc ttaatgtatg ttaaattgaa gaaggtttgt
4261 ataccaacag atattttttt gaatctaaga aaagctcaac aaacttttgg ggaaaatgaa
4321 actgccatag gacttttgac caaaggcttg acgacaaaca cataccctgt tagcatgaat
4381 tggttgcaag gcaatttaaa ttatctgtct tctgtttatc actcttgtgc aatgaaagct
4441 tatcacaaca ctttggaatg ttacaaaaac tgtgatttcc aaactagatg gattgtgcac

4501 tctgatgata atgcaacatc attaatagcc agtggagagg ttgataaaat gctgacagac
4561 ttttcaagct catctctgcc agaaatgttg tttagaagca ttgaagctca tttcaaaagt
4621 ttttgcataa ctttgaaccc aaaaaagagt tatgcttctt catcagaagt agagtttata
4681 tctgaaagaa ttagtaaatg gagcgattat tcctctctat tgcaggcatt tagcaaactg
4741 ttgcacagaa tcttcgcata taagttattt gatgatctaa tgtcactcag tatacatgtt
4801 acaatgcttc tgagaaaagg ctgtcctaat gaagttatac cttttgctta tggggctgtg
4861 caggtacaag cgttaagcat ctattcaatg cttcctggtg aagtgaatga tagtattaga
4921 atttttaaca agcttggagt aagtttaaag tcaaacgaga ttcccacaaa catggggggc
4981 tggttgacct ctcctataga gccgttgtct atattaggtc catcatcaaa tgatcaaatc
5041 atctattaca atgtgataag agattttttg aacaaaaaaa gtttagaaga agtaaaagat
5101 agtgtctctt cttccagtta tctacagatg agattcagag agctaaaaga aaagtatgaa
5161 agaggaactc tggaagaaaa agataaaaag atgatatttc ttatcaatct gtttgagaaa
5221 gcatcagtgt ctgaagattc agacgttcta acaattggga tgaaatttca aactatgtta
5281 actcagatta taaaattacc taattttata aatgagaatg ctttaaacaa gatgtcaagt
5341 tataaagatt tttcaaaact ttatcctaat ttaaaaaaga atgaagattt atataaaagc
5401 actaagaact taaagataga cgaggatgct gttttagagg aagatgagtt atataagaag
5461 attgcatcta gcttagaaat ggaatctgtc catgacataa tgataaaaaa tcctgaaaca
5521 attctgatag caccattgaa tgatagagat tttttactta gtcagctgtt catgtacaca
5581 agcccttcta aaagaaacca gttatcgaac caatctacag agaaacttgc tttagataga
5641 gtgttaaggt caaaagctag aacatttgta aacatttctt ccactgtgaa gatgacttat
5701 gaagaaaaca tggaaaagaa aatcttagaa atgctaaaat ttgatttaga ttcatattgt
5761 tcatttaaaa catgtgtaaa tctagttatc aaggatgtta atttcagcat gctgattcca
5821 atattagatt ctgcataccc ttgtgaatct aggaaaagag ataactacaa tttcaggtgg
5881 tttcagactg agagatggat acctgttgtt gaaggctctc cgggactagt agtaatgcat
5941 gctgtctatg gatcaaatta tatagagaac ttaggtttaa aaaacatccc tctaacagac
6001 gatagtatta atgttttaac aagcacgttt ggaacaggtt taatcatgga agatgtaaaa


6061 tccctagtta atggcaaaga cagcttcgaa acagaggctt ttagcaattc taatgaatgt
6121 caaagattgg tgaaagcatg caattatatg atagcagcac aaaacaggct tttagcaatt

6181 aacacatgct ttactaggaa aagctttccc ttctattcta agttcaatct agggagaggg
6241 tttatctcaa acacattagc tctcctatcc accatctaca gtaaagaaga atcctatcat
6301 tttgtttcta cagctagtta taaattagac aaaactatca gaactgtggt aagtgctcag
6361 caagatatga acttagagaa aatactggac actgctgtat acatatcaga taaattgcag
6421 tcacttttcc caacaattac aagagaggat atagttttaa tattgcaaaa tgtttgcctt
6481 gacagtaaac ctatatggca gagtctagaa gacaaaatga aaaagattaa caattcaaca
6541 gcaagtggct tcacagtgtc aaatgtgatt ctatcacata acagtgaatt gaacacaatc
6601 cagaaacaaa ttgtctggat gtggaacatg ggtttgtgtt ctcacagaac attagatttt
6661 gttatcaggt atattagaag aagggatgta agatatgtaa aaactgaaga acaagatgaa
6721 tcaggaaatt atgtctctgg aactatgtac aaaataggga tcatgacaag aagctgctat
6781 gtggaattga tagcatctga tcaagatgta gcagtttctt tgagaacacc atttgagata
6841 ttgaatgaaa gagagtatct ttttgacaca tacagagaaa gtatagagaa attattggca
6901 gaaattatgt ttgataaagt gaacataata aatcaaacaa ccacagattg ttttcttaga
6961 accaggagat cttgcatcag aatgaccaca gacaacaaaa tgattgtaaa ggttaatgct
7021 acatcaagac aaataagact agagaatgta aaattagttg taaagataaa atatgaaaat
7081 gtgaattccg atgtatggga tattatagaa agccaaaaat ctctagtctt aaggctccct
7141 gaagtagggg aatttttctc tgatatgtat aaaactgcag actctgaaac tgaaacaatc
7201 aaaaccataa aaaacaggct tatgacttct ttaactttca tagaagcctt tggaaactta
7261 tcacagcaga tcaaagagat tgtagatgat gatatcagag aaacgatgga tgaattctta
7321 atgaacatcc gggatacctg cttagaaggt ttggaaaact gcaaaagtgt ggaagaatat
7381 gatagctatc ttgatgaaaa tggatttaat gacacagtag aactattcga aaacttgcta
7441 agaacacatg acaactttga aaatgagtat agtcctcttt tttcagagat tgttgacaaa
7501 gcaaaacagt atactagaga tttagaaggt ttcaaagaaa tactgctcat gcttaaatat
7561 tctctaataa atgatgcatc aggatttaaa agctatagag ccactggaat gcatgctgtt
7621 gagctaatgg caaaaaagca catagagata ggggaattca acttgttagg aatgatccaa
7681 ttgattaaag cttgtgaaac atgccacaac aatgactcta tattaaactt agcaagttta
7741 aggaatgttc ttagcaggac atatgccaca tttgggagga gaataagatt ggatcatgat
7801 ctggacttgc aaaacaactt aatggaaaaa agttatgatt tcaagacgct ggttttacca
7861 gaaataaaat tatcagaact atctagggaa atactgaaag aaaatgggtt tgttatatct
7921 ggagagaatc taaaaatgga taggtctgat gaagaatttg tgggtcttgc cagttttaat

7981 gtgttgaggc tagatgagga agaaatgtat gaaggtttga tcaaagaaat gaaaattaaa
8041 aggaaaaaga aagggttttt atttccagca aacacacttc tactaagtga gttgataaag
8101 ttcttgattg gaggaataaa gggaaccagc tttgatatag agacattgtt acggaacagt
8161 tttagaccag acatattttc aactgacaga ttgggaagat taagttccag tgtacctgca
8221 ctcaaagttt atgcaactgt ttatatggaa tataagaatg tcaattgtcc tttaaatgag
8281 atagctgaca gcttagaagg ttatctaaaa ctgacaaaaa gcaggtccaa ggaacatttc
8341 ttgtctggaa gagttaaaaa agctttgata caattaagag atgaacaatc gcgaactaaa
8401 aaactagagg tctataagga tatcgcaaat ttccttgcta ggcacccact atgtttatca
8461 gaaaaaacat tgtatggaag atatacctac tctgatatca atgattatat catgcaaaca
8521 agagagatta ttttgagtaa aataagtgag ttggacgagg ttgttgaaac agatgaagac
8581 aatttcttgc ttagttatct aagaggggaa gaagatgcct ttgatgaaga tgagcttgat
8641 gaagaagaag acacagatta aattgaaagt aatgactaac aatccatgaa taacagatta
8701 gatataactt agaatataaa tttattgcta ttttagaatt agattagatc tacttagcct
8761 aaaacaattt ggtgaaccaa atctatagtg tatataaatg tagagtcccg gtatagtttc
8821 actggaggga attcttatgt aatttgtaaa gtctggctgt ggagaggtta tatgttttag
8881 ttgtacctga ttgctct
Hình pl.1c: Trình tự Nucleotide genome L (Long)