Luận văn : SỬ DỤNG KỸ THUẬT DAS-ELISA VÀ RT-PCR ĐỂ PHÁT HIỆN VIRUS GÂY BỆNH ĐỐM VÕNG TRÊN CÂY ĐU ĐỦ (Papaya ringspot virus) TẠI HAI TỈNH ĐỒNG NAI VÀ ĐỒNG THÁP part 9 ppsx

69


Kết quả đọc trình tự của mồi M31 trên mẫu Mỹ Hiệp




70


Sự tƣơng đồng giữa hai mồi P14 và M31 của mẫu Mỹ Hiệp



CLUSTAL X (1.83) multiple sequence alignment

25
ABI3100_Ic-P14_008_2005-08-2 1 CTTGCTTGATCGCTATGTCGATAG AAAAGATAACAAAAAGAAGAAAAAGA 50
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 51 TAAACAAAAAGACAAGAATTCATGAAGGTACTAGTGACGGAAACAATGTG 100
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 101 TCAACTAGTACCAAAACTGGAGAGAGAGATAGAGATGTCAATGCCGGAAC 150
ABI3100_Ic-M31_010_2005-08-2

ABI3100_Ic-P14_008_2005-08-2 151 TAGTGGAACTTTCACTGTTCCGAGAATAAAATCATTTACTGACAAGATCA 200
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 201 TTTTACCGAAAATTAAAGGAAAAACTGTCCTTAATTTAAATCATCTTCTT 250
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 251 CAGTATAATCCGCAACAAATTGATATCTCAAACACTCGTGCCACTCAAAC 300
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 301 TCAATTCGAAAAGTGGTATGGGGGAGTGAGGAATGATTACGGTCTCAACG 350
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 351 ATAACGAAATGCAAGTGATGTTAAACGGTTTGATGGTTTGGTGTATTGAA 400
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 401 AATGGTACATCTCCAGACTTATCTGGTGTTTGGGTAATGATGGATGGGGA 450
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 451 AACACAAGTTGATTATCCCATTAAGCCTTTAATTGAACATGCAACTCCTT 500
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 501 CGTTTAGGCAAATCATGGCTCACTTCAGTAACGCGGCAGAGGCATACATC 550

ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 551 GCGAAGAGGAATGCAACTGAAAGGTATATGCCGCGGTATGGAATCAAGAG 600
ABI3100_Ic-M31_010_2005-08-2


ABI3100_Ic-P14_008_2005-08-2 601 GAATTTGACTGACATTAGCCTCGCTAGATACGCTTTCG ATA 641
ABI3100_Ic-M31_010_2005-08-2 NNNNN
639


71


Phụ lục 9

Kết quả giải trình tự với mồi P14 của mẫu Định Quán




72


Kết quả giải trình từ với mồi M30 của Định Quán

73


Sự tƣơng đồng của hai mồi P7 và M30 của mẫu Định Quán


CLUSTAL X (1.83) multiple sequence alignment


ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 1 CCTCCCCGCGGCTCTCTCCCGCGAAAAATCTCCCGTTTTAATGGCCCTTT 50


ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 51 TGGGGGAGGGTGTTAATAATATGTTTTTTTTTTTGTGGGGGGCCCCCTCC 100


ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 101 CCCCCCACACACCACTATTTTTTTTTTTTAACCCACAAAAAAAAAACAAC 150


ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 151 CTAAAACCCCTCTTTTTGTGGTGGAGAGAATAGACCCCCCCCCCCCTAAA 200


ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 201 AAATATAAAAAAAGTGATGCGCAGTTGGGGTTACCAATTGGGCTTTGAAG 250

ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 251 AAAAATTTTTTAATATTAACCTTTTCCCCTTATTTTTTTAATATCATCTT 300

341-m30
ABI3100_DQ17-P7_015_2005-08-
ABI3100_DQ17-M30_002_2005-08 301 GTCAGTAAATGATTTTATTCTAATCTCGGGACCAGTGGAA CTTTCACGGT 350

1
ABI3100_DQ17-P7_015_2005-08- CGAGAATAAAATCATTTACTGACAAGATGATATTACCAAAAATTAAGG 47
ABI3100_DQ17-M30_002_2005-08 351 TCCGAGAATAAAATCATTTACTGACAAGATGATATTACCAAAAATTAAGG 400
* ** *************** ***********************

ABI3100_DQ17-P7_015_2005-08- 48 GAAAAACTGTCCTTAATTTAAATCATCTTCTTCAGTATAATCCGCAACAA 97
ABI3100_DQ17-M30_002_2005-08 401 GAAAAACTGTCCTTAATTTAAATCATCTTCTTCAGTATAATCCGCAACAA 450
**************************************************

ABI3100_DQ17-P7_015_2005-08- 98 ATTGATATCTCAAACACTCGTGCCACTCAAACTCAATTTGAAAAGTGGTA 147
ABI3100_DQ17-M30_002_2005-08 451 ATTGATATCTCAAACACTCGTGCCACTCAAACTCAATTTGAAAAGTGGTA 500
**************************************************

ABI3100_DQ17-P7_015_2005-08- 148 TGAGGGAGTGCGGAATGATTACGGTCTTAACGATGACGAAATGCAAGTGA 197
ABI3100_DQ17-M30_002_2005-08 501 TGAGGGAGTGCGGAATGATTACGGTCTTAACGATGACGAAATGCAAGTGA 550
**************************************************

ABI3100_DQ17-P7_015_2005-08- 198 TGTTAAATGGTTTGATGGTTTGGTGTATCGAAAATGGTACATCTCCAGAC 247
ABI3100_DQ17-M30_002_2005-08 551 TGTTAAATGGTTTGATGGTTTGGTGTATCGAAAATGGTACATCTCCAGAC 600
**************************************************

ABI3100_DQ17-P7_015_2005-08- 248 TTGTCTGGTGTTTGGGTAATGATGGATGGGGAAATACAAGTAGATTATCC 297

ABI3100_DQ17-M30_002_2005-08 601 TTGTCTGGTGTTTGGGTAATGATGGATGGGGAAATACAAGTAGATTATCC 649
*********************** * * **********************

ABI3100_DQ17-P7_015_2005-08- 298 CATCA-AGCCTTTAATTGAACATGCAACTCCCTCGTTTAGGCAAATCATG 346
ABI3100_DQ17-M30_002_2005-08 650 CATCA-AGCCTTTAATTGAACATGCAACTCCCTCGTTTAGGCAAATCATG 699
**** ************* ************************ *****

ABI3100_DQ17-P7_015_2005-08- 347 GCTCACTTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGAATG CCACT 396
ABI3100_DQ17-M30_002_2005-08 701 GCTCACTTCAGTAACGCGGCAGAGGCATACAT
****** ******************* * *732 391-P7

ABI3100_DQ17-P7_015_2005-08- 397 GAGAGGAGTGAAAGTTCCCCTGGTCCCCAAAACGGAGGGGTAGGAAAAGG 446
ABI3100_DQ17-M30_002_2005-08


ABI3100_DQ17-P7_015_2005-08- 447 A
ABI3100_DQ17-M30_002_2005-08 -



74


Phụ lục 10

Kết quả giải trình tự với mồi P7 của mẫu Mỹ Hiệp

75



Kết quả giải trình tự với mồi M30 của mẫu Mỹ Hiệp





76



Sự tƣơng đồng của hai mồi P7 và M30 của mẫu Mỹ Hiệp


CLUSTAL X (1.83) multiple sequence alignment


ABI3100_I-P7_004_2005-08-24
ABI3100_I-M30_006_2005-08-24 1 TGATTTGTAATTACACCACGTTTTCGCCTTCAAACTTTTCTTGGATATGC 50

ABI3100_I-P7_004_2005-08-24
ABI3100_I-M30_006_2005-08-24 51 TCAACAAACTTCTCTGTCCGTGAACAATGGAGCAGTCAAGCAATTGGCGG 100


ABI3100_I-P7_004_2005-08-24
ABI3100_I-M30_006_2005-08-24 101 ATAAGAGGTTTAGGGGTGTGAGTGGGTTTTTCCGGTCAACTCTCTCGGGG 150

1
ABI3100_I-P7_004_2005-08-24 CGATCGA 7
ABI3100_I-M30_006_2005-08-24 151 AGATTCATGTGCTAAATTCGGACCCAGTGGAACTTTCACGGTTCGATCGA 200
* *

ABI3100_I-P7_004_2005-08-24 8 GATAATCATTTACTGACA-GATGATTTTACCGAAAATTAAAGGAAAAACT 56
ABI3100_I-M30_006_2005-08-24 201 GATAATCATTTACTGACA-GATGATTTTACCGAAAATTAAAGGAAAAACT 250
* **** ********** ************* *****************

ABI3100_I-P7_004_2005-08-24 57 GTCCTTAATTTTAAATCATCTTCTTCAGTATAATCCGCAACAAATTGATA 106
ABI3100_I-M30_006_2005-08-24 251 GTCCTTAATTTTAAATCATCTTCTTCAGTATAATCCGCAACAAATTGGTA 299
**** ****** *********** ********** ********* **

ABI3100_I-P7_004_2005-08-24 107 TCTCAAACACTCGTGCCACTCAAACTCAATTCGAAAAGTG-GTATGAGGG 155
ABI3100_I-M30_006_2005-08-24 300 TCTCAAACACTCGTGCCTCTCAAACTCAATTCGAAAAGTG-GTATGAGGG 349
******* ** ***** ******* *** **** * **

ABI3100_I-P7_004_2005-08-24 156 AGTGAGGAATGATTACGGTCTCAACGATAACGAAATGCAAGTG-ATGTTA 204
ABI3100_I-M30_006_2005-08-24 350 AGTGAGGAATGATTACGGTCTCAACGATAACGAAATGCAAGTG-ATGTTA 399
** *** *** ** * * ** * ***** ***** * * *

ABI3100_I-P7_004_2005-08-24 205 AACGGTTTGATGGTTTGGTGTATTGAAAATGGTACATCTCCAGACTTATC 254
ABI3100_I-M30_006_2005-08-24 400 AACGGTTTGATGGTTTGGTGTATTGAAAATGGTACATCTCCAGACTTATC 449
** * * ********* * * ****** ****** ****** ***

ABI3100_I-P7_004_2005-08-24 255 TGGTGTTTGGGTAATGATGGATGGGGAAACACAAGTTGATTATCCCATTA 304
ABI3100_I-M30_006_2005-08-24 450 TGGTGTTTGGGTAATGATGGATGGGGAAACACAAGTTGATTATCCCATTA 499
* ******************** ************** ****** ***

ABI3100_I-P7_004_2005-08-24 305 AGCCTTTAATTGAACATGC AACTCCTTCGTTTAGGCAAATCATGGCTCAC 354

ABI3100_I-M30_006_2005-08-24 500 AGCCTTTAATTGAACATGC AACTCCTTCGTCCAGGCAAATCATGGCTCAC 544
******************* ** * ** ** * * *

ABI3100_I-P7_004_2005-08-24 255 TTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGAAATGCCACTGAAAG 404
ABI3100_I-M30_006_2005-08-24 545 TTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGAAATGCCACTGAAAG 589
*** * * * * *** * * **** * * ** ****

ABI3100_I-P7_004_2005-08-24 405 GAATATGCCGCGGTATGGAATCAAGAGGAATTTGACTGACATTAGTCT 452
ABI3100_I-M30_006_2005-08-24 590 GAATATGCCGCGGTATGGAATCAAGAGGAATTTGACTGACATTAGTCT 639
** * ** *** * * * ** ** * *

ABI3100_I-P7_004_2005-08-24 453 CGCCAGAT ATGCTTTCGACTTCTATG AGGTGAATTCAAA AACAC 496
ABI3100_I-M30_006_2005-08-24 640 CGCCAGAT ATGCTTTCGACTTCTATG AGGTGAATTCAAA AACAC 689
* ** ** ** * ** ** * ** **** * *

ABI3100_I-P7_004_2005-08-24 497 CTGATA-GAGCTCGTGAAGCTCATATGCAGA-TGAAGGCTGCAGCACTGC 544
ABI3100_I-M30_006_2005-08-24 690 GTGATA-GAGCTCGTGAAGCTCATATGCAGA-TGAAGGCTGCAGCACTGC 739
** ** *** **** * * * * * ** * * * *** ***

ABI3100_I-P7_004_2005-08-24 545 GTACTACTGGTCGCAGAATGTTTGGAATGGACGGCAGTGTCAGTAACAAG 594
ABI3100_I-M30_006_2005-08-24 740 GTACTACTGGTCGCAGAATGTTTGGAATGGACGGCAGTGTCAGTAACAAG 789
* * ** * *** * *** ** *** * *** * *

ABI3100_I-P7_004_2005-08-24 595 GAAGAAAACACGGAGAGACACACAGTGGAAGATGTCAATAGA-GACATGC 643
ABI3100_I-M30_006_2005-08-24 790 GAAGAAAACACGGAGAGACACACAGTGGAAGATGTCAATAGA-GACATGC 839
* ******* *** *** * * * * * * * * ****

ABI3100_I-P7_004_2005-08-24 644 ACTCTCTCCTGGGTATGCGCAACTGA-ATACTCGCCCATGGTTAGCTAAC 692
ABI3100_I-M30_006_2005-08-24 840 ACTCTCTCCTGGGTATGCGCAACTGA-ATACTCGCCCATGGTTAGCTAAC 899
* * ** * ***** * ** * * *** ************

77





ABI3100_I-P7_004_2005-08-24 693 CATGGGAGAGCACTCAGTTGCGCGGCAGAGGAAGAGAGCGCATGCACAAG 730
ABI3100_I-M30_006_2005-08-24 890 CATGGGAGAGCACTCAGTTGCGCGGCAGAGGAAGAGAG 935
****** *** * ***** ************ * *926

ABI3100_I-P7_004_2005-08-24 743 TGCCCCTGAAAGACTGTGCCCTCTCCGTGGTTTCATACAAGAAAGAGACA 792
ABI3100_I-M30_006_2005-08-24


ABI3100_I-P7_004_2005-08-24 793 AAGGCGTCCCTTCAAAAATTCTGCGACTGTATTACGCGAGCTGGAGCTTT 842
ABI3100_I-M30_006_2005-08-24


ABI3100_I-P7_004_2005-08-24 843 CTAAGGAATATGGCTCCCGAGCTAAATAGAG 873
ABI3100_I-M30_006_2005-08-24



78


Phụ lục 11
Đoạn tƣơng đồng chung cho cả 3 mẫu

CLUSTAL X (1.83) multiple sequence alignment

DQ
Ic 1 AAAAGATAACAAAAAGAAGAAAAAGATAAACAAAAAGACAAGAATTCATGAAGGTACTAG 60
I


DQ
Ic 61 TGACGGAAACAATGTGTCAACTAGTACCAAAACTGGAGAGAGAGATAGAGATGTCAATGC 120
I

1 12
DQ 1 CTTTCACGGTT CCGAGAATAAAATCATTTACTGACAAGATGATTTT 46
Ic 121 CGGAACTAGTGGAACTTTCACTGTT CCGAGAATAAAATCATTTACTGACAAGATGATTTT 180
I CCGAGAATAAAATCATTTACTGACAAGATGATTTT 34
1* * * *************** *** ** **

DQ 47 ACCGAAAATTAAAGGAAAAACTGTCCTTAATTTTAAATCATCTTCTTCAGTATAATCCGC 105
Ic 181 ACCGAAAATTAAAGGAAAAACTGTCCTTAATTTTAAATCATCTTCTTCAGTATAATCCGC 239
I 35 ACCGAAAATTAAAGGAAAAACTGTCCTTAATTTTAAATCATCTTCTTCAGTATAATCCGC 94
*** ******** ******************** **************************

DQ 106 AACAAATTGATATCTCAAACACTCGTGCCACTCAAACTCAATTCGAAAAGTG-GTATGAG 164
Ic 240 AACAAATTGATATCTCAAACACTCGTGCCACTCAAACTCAATTCGAAAAGTG-GTATGAG 298
I 95 AACAAATTGATATCTCAAACACTCGTGCCACTCAAACTCAATTCGAAAAGTG-GTATGAG 153
******************************************* ******** ***** *

DQ 165 GGAGTGAGGAATGATTACGGTCTCAACGATAACGAAATGCAAGTG-ATGTTAAACGGTTT 223
Ic 299 GGAGTGAGGAATGATTACGGTCTCAACGATAACGAAATGCAAGTG-ATGTTAAACGGTTT 357

I 154 GGAGTGAGGAATGATTACGGTCTCAACGATAACGAAATGCAAGTG-ATGTTAAACGGTTT 212
****** **************** ****** ************** ******** *****

DQ 224 GATGGTTTGGTGTATTGAAAATGGTACATCTCCAGACTTATCTGGTGTTTGGGTAATGAT 283
Ic 358 GATGGTTTGGTGTATTGAAAATGGTACATCTCCAGACTTATCTGGTGTTTGGGTAATGAT 417
I 213 GATGGTTTGGTGTATTGAAAATGGTACATCTCCAGACTTATCTGGTGTTTGGGTAATGAT 272
*************** *********************** ********************

DQ 284 GGATGGGGAAACACAAGTTGATTATCCCATTAAGCCTTTAATTGAACATGCAACTCCTTC 343
Ic 418 GGATGGGGAAACACAAGTTGATTATCCCATTAAGCCTTTAATTGAACATGCAACTCCTTC 477
I 273 GGATGGGGAAACACAAGTTGATTATCCCATTAAGCCTTTAATTGAACATGCAACTCCTTC 332
*********** ****** *********** ************************** **

DQ 344 GTTTAGGCAAATCATGGCTCACTTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGGAA 402
Ic 478 GTTTAGGCAAATCATGGCTCACTTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGGAA 537
I 333 GTTTAGGCAAATCATGGCTCACTTCAGTAACGCGGCAGAGGCATACATCGCAAAGAGGAA 392
*************************************************** ***** **
404
DQ 403 TG
Ic 538 TG CAACTGAAAG GTATATGCCGCGGTATGGAATCAAGAGGAATTTGACTGACATTAGC 595
I 393 TG CCACTGAAAG GAATATGCCGCGGTATGGAATCAAGAGGAATTTGACTGACATTAGT 450
**

DQ
Ic 596 CTCGCTAGAT ACGCTTTCG 614
I 451 CTCGCCAGAT ATGCTTTCGACTTCTATG AGGTGAATTCAAA AACACCTGATA-G 503

DQ
Ic
I 504 AGCTCGTGAAGCTCATATGCAGA-TGAAGGCTGCAGCACTGCGTACTACTGGTCGCAGAA 562

DQ
Ic
I 563 TGTTTGGAATGGACGGCAGTGTCAGTAACAAGGAAGAAAACACGGAGAGACACACAGTGG 622

DQ
Ic
I 623 AAGATGTCAATAGA-GACATGCACTCTCTCCTGGGTATGCGCAACTGA-ATACTCGCCCA 680

DQ
Ic
I 681 TGGTTAGCTAACCATGGGAGAGCACTCAGTTGCGCGGCAGAGGAAGAGAG 730

Đoan chung cho ca ba = 393