﻿                            序列表

<110>  江苏省农业科学院

<120>  基于基因编辑技术的ALS突变型蛋白及其基因在植物育种中的应用  

<160>  31

<170>  SIPOSequenceListing 1.0

<210>  1
<211>  1935
<212>  DNA
<213>  ALS-nj突变型基因(acetolactate synthase)

<400>  1
atggctacga ccgccgcggc cgcggccgcc gccctgtccg ccgccgcgac ggccaagacc        60
ggccgtaaga accaccagcg acaccacgtc cttcccgctc gaggccgggt gggggcggcg       120
gcggtcaggt gctcggcggt gtccccggtc accccgccgt ccccggcgcc gccggccacg       180
ccgctccggc cgtgggggcc ggccgagccc cgcaagggcg cggacatcct cgtggaggcg       240
ctggagcggt gcggcgtcag cgacgtgttc gcctacccgg gcggcgcgtc catggagatc       300
caccaggcgc tgacgcgctc cccggtcatc accaaccacc tcttccgcca cgagcagggc       360
gaggcgttcg cggcgtccgg gtacgcgcgc gcgtccggcc gcgtcggggt ctgcgtcgcc       420
acctccggcc ccggggcaac caacctcgtg tccgcgctcg ccgacgcgct gctcgactcc       480
gtcccgatgg tcgccatcac gggccaggtc ccccgccgca tgatcggcac cgacgccttc       540
caggagacgc ccatagtcga ggtcacccgc tccatcacca agcacaatta ccttgtcctt       600
gatgtggagg acatcccccg cgtcatacag gaagccttct tcctcgcgtc ctcgggccgt       660
cctggcccgg tgctggtcga catccccaag gacatccagc agcagatggc cgtgccggtc       720
tgggacacct cgatgaatct accagggtac atcgcacgcc tgcccaagcc acccgcgaca       780
gaattgcttg agcaggtctt gcgtctggtt ggcgagtcac ggcgcccgat tctctatgtc       840
ggtggtggct gctctgcatc tggtgacgaa ttgcgctggt ttgttgagct gactggtatc       900
ccagttacaa ccactctgat gggcctcggc aatttcccca gtgacgaccc gttgtccctg       960
cgcatgcttg ggatgcatgg cacggtgtac gcaaattatg ccgtggataa ggctgacctg      1020
ttgcttgcgt ttggtgtgcg gtttgatgat cgtgtgacag ggaaaattga ggcttttgca      1080
agcagggcca agattgtgca cattgacatt gatccagcag agattggaaa gaacaagcaa      1140
ccacatgtgt caatttgcgc agatgttaag cttgctttac agggcttgaa tgctctgcta      1200
caacagagca caacaaagac aagttctgat tttagtgcat ggcacaatga gttggaccag      1260
cagaagaggg agtttcctct ggggtacaaa acttttggtg aagagatccc accgcaatat      1320
gccattcagg tgctggatga gctgacgaaa ggtgaggcaa tcatcgctac tggtgttggg      1380
cagcaccaga tgtgggcggc acaatattac acctacaagc ggccacggca gtggctgtct      1440
tcggctggtc tgggcgcaat gggatttggg ctgcctgctg cagctggtgc ttctgtggct      1500
aacccaggtg tcacagttgt tgatattgat ggggatggta gcttcctcat gaacattcag      1560
gagctggcat tgatccgcat tgagaacctc cctgtgaagg tgatggtgtt gaacaaccaa      1620
catttgggta tggtggtgca atgggaggat aggttttaca aggcgaatag ggcgcataca      1680
tacttgggca acccggaatg tgagagcgag atatatccag attttgtgac tattgctaag      1740
gggttcaata ttcctgcagt ccgtgtaaca aagaagagtg aagtccgtgc cgccatcaag      1800
aagatgctcg agactccagg gccatacttg ttggatatca tcgtcccgca ccaggagcat      1860
gtgctgccta tgatcccaag ttggggcgca ttcaaggaca tgatcctgga tggtgatggc      1920
aggactgtgt attaa                                                       1935

<210>  2
<211>  644
<212>  PRT
<213>  ALS突变型蛋白(acetolactate synthase)

<400>  2
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ser Ala Ala Ala 
1               5                   10                  15      
Thr Ala Lys Thr Gly Arg Lys Asn His Gln Arg His His Val Leu Pro 
            20                  25                  30          
Ala Arg Gly Arg Val Gly Ala Ala Ala Val Arg Cys Ser Ala Val Ser 
        35                  40                  45              
Pro Val Thr Pro Pro Ser Pro Ala Pro Pro Ala Thr Pro Leu Arg Pro 
    50                  55                  60                  
Trp Gly Pro Ala Glu Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala 
65                  70                  75                  80  
Leu Glu Arg Cys Gly Val Ser Asp Val Phe Ala Tyr Pro Gly Gly Ala 
                85                  90                  95      
Ser Met Glu Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Thr Asn 
            100                 105                 110         
His Leu Phe Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr 
        115                 120                 125             
Ala Arg Ala Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro 
    130                 135                 140                 
Gly Ala Thr Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser 
145                 150                 155                 160 
Val Pro Met Val Ala Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly 
                165                 170                 175     
Thr Asp Ala Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile 
            180                 185                 190         
Thr Lys His Asn Tyr Leu Val Leu Asp Val Glu Asp Ile Pro Arg Val 
        195                 200                 205             
Ile Gln Glu Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val 
    210                 215                 220                 
Leu Val Asp Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val 
225                 230                 235                 240 
Trp Asp Thr Ser Met Asn Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys 
                245                 250                 255     
Pro Pro Ala Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu 
            260                 265                 270         
Ser Arg Arg Pro Ile Leu Tyr Val Gly Gly Gly Cys Ser Ala Ser Gly 
        275                 280                 285             
Asp Glu Leu Arg Trp Phe Val Glu Leu Thr Gly Ile Pro Val Thr Thr 
    290                 295                 300                 
Thr Leu Met Gly Leu Gly Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu 
305                 310                 315                 320 
Arg Met Leu Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp 
                325                 330                 335     
Lys Ala Asp Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val 
            340                 345                 350         
Thr Gly Lys Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile 
        355                 360                 365             
Asp Ile Asp Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser 
    370                 375                 380                 
Ile Cys Ala Asp Val Lys Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu 
385                 390                 395                 400 
Gln Gln Ser Thr Thr Lys Thr Ser Ser Asp Phe Ser Ala Trp His Asn 
                405                 410                 415     
Glu Leu Asp Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe 
            420                 425                 430         
Gly Glu Glu Ile Pro Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu 
        435                 440                 445             
Thr Lys Gly Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met 
    450                 455                 460                 
Trp Ala Ala Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser 
465                 470                 475                 480 
Ser Ala Gly Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly 
                485                 490                 495     
Ala Ser Val Ala Asn Pro Gly Val Thr Val Val Asp Ile Asp Gly Asp 
            500                 505                 510         
Gly Ser Phe Leu Met Asn Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu 
        515                 520                 525             
Asn Leu Pro Val Lys Val Met Val Leu Asn Asn Gln His Leu Gly Met 
    530                 535                 540                 
Val Val Gln Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr 
545                 550                 555                 560 
Tyr Leu Gly Asn Pro Glu Cys Glu Ser Glu Ile Tyr Pro Asp Phe Val 
                565                 570                 575     
Thr Ile Ala Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys 
            580                 585                 590         
Ser Glu Val Arg Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro 
        595                 600                 605             
Tyr Leu Leu Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met 
    610                 615                 620                 
Ile Pro Ser Trp Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly 
625                 630                 635                 640 
Arg Thr Val Tyr 
                

<210>  3
<211>  1935
<212>  DNA
<213>  ALS野生型基因(acetolactate synthase)

<400>  3
atggctacga ccgccgcggc cgcggccgcc gccctgtccg ccgccgcgac ggccaagacc        60
ggccgtaaga accaccagcg acaccacgtc cttcccgctc gaggccgggt gggggcggcg       120
gcggtcaggt gctcggcggt gtccccggtc accccgccgt ccccggcgcc gccggccacg       180
ccgctccggc cgtgggggcc ggccgagccc cgcaagggcg cggacatcct cgtggaggcg       240
ctggagcggt gcggcgtcag cgacgtgttc gcctacccgg gcggcgcgtc catggagatc       300
caccaggcgc tgacgcgctc cccggtcatc accaaccacc tcttccgcca cgagcagggc       360
gaggcgttcg cggcgtccgg gtacgcgcgc gcgtccggcc gcgtcggggt ctgcgtcgcc       420
acctccggcc ccggggcaac caacctcgtg tccgcgctcg ccgacgcgct gctcgactcc       480
gtcccgatgg tcgccatcac gggccaggtc ccccgccgca tgatcggcac cgacgccttc       540
caggagacgc ccatagtcga ggtcacccgc tccatcacca agcacaatta ccttgtcctt       600
gatgtggagg acatcccccg cgtcatacag gaagccttct tcctcgcgtc ctcgggccgt       660
cctggcccgg tgctggtcga catccccaag gacatccagc agcagatggc cgtgccggtc       720
tgggacacct cgatgaatct accagggtac atcgcacgcc tgcccaagcc acccgcgaca       780
gaattgcttg agcaggtctt gcgtctggtt ggcgagtcac ggcgcccgat tctctatgtc       840
ggtggtggct gctctgcatc tggtgacgaa ttgcgctggt ttgttgagct gactggtatc       900
ccagttacaa ccactctgat gggcctcggc aatttcccca gtgacgaccc gttgtccctg       960
cgcatgcttg ggatgcatgg cacggtgtac gcaaattatg ccgtggataa ggctgacctg      1020
ttgcttgcgt ttggtgtgcg gtttgatgat cgtgtgacag ggaaaattga ggcttttgca      1080
agcagggcca agattgtgca cattgacatt gatccagcag agattggaaa gaacaagcaa      1140
ccacatgtgt caatttgcgc agatgttaag cttgctttac agggcttgaa tgctctgcta      1200
caacagagca caacaaagac aagttctgat tttagtgcat ggcacaatga gttggaccag      1260
cagaagaggg agtttcctct ggggtacaaa acttttggtg aagagatccc accgcaatat      1320
gccattcagg tgctggatga gctgacgaaa ggtgaggcaa tcatcgctac tggtgttggg      1380
cagcaccaga tgtgggcggc acaatattac acctacaagc ggccacggca gtggctgtct      1440
tcggctggtc tgggcgcaat gggatttggg ctgcctgctg cagctggtgc ttctgtggct      1500
aacccaggtg tcacagttgt tgatattgat ggggatggta gcttcctcat gaacattcag      1560
gagctggcat tgatccgcat tgagaacctc cctgtgaagg tgatggtgtt gaacaaccaa      1620
catttgggta tggtggtgca atgggaggat aggttttaca aggcgaatag ggcgcataca      1680
tacttgggca acccggaatg tgagagcgag atatatccag attttgtgac tattgctaag      1740
gggttcaata ttcctgcagt ccgtgtaaca aagaagagtg aagtccgtgc cgccatcaag      1800
aagatgctcg agactccagg gccatacttg ttggatatca tcgtcccgca ccaggagcat      1860
gtgctgccta tgatcccaag tgggggcgca ttcaaggaca tgatcctgga tggtgatggc      1920
aggactgtgt attaa                                                       1935

<210>  4
<211>  644
<212>  PRT
<213>  ALS野生型蛋白(acetolactate synthase)

<400>  4
Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala Leu Ser Ala Ala Ala 
1               5                   10                  15      
Thr Ala Lys Thr Gly Arg Lys Asn His Gln Arg His His Val Leu Pro 
            20                  25                  30          
Ala Arg Gly Arg Val Gly Ala Ala Ala Val Arg Cys Ser Ala Val Ser 
        35                  40                  45              
Pro Val Thr Pro Pro Ser Pro Ala Pro Pro Ala Thr Pro Leu Arg Pro 
    50                  55                  60                  
Trp Gly Pro Ala Glu Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala 
65                  70                  75                  80  
Leu Glu Arg Cys Gly Val Ser Asp Val Phe Ala Tyr Pro Gly Gly Ala 
                85                  90                  95      
Ser Met Glu Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Thr Asn 
            100                 105                 110         
His Leu Phe Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr 
        115                 120                 125             
Ala Arg Ala Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser Gly Pro 
    130                 135                 140                 
Gly Ala Thr Asn Leu Val Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser 
145                 150                 155                 160 
Val Pro Met Val Ala Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly 
                165                 170                 175     
Thr Asp Ala Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile 
            180                 185                 190         
Thr Lys His Asn Tyr Leu Val Leu Asp Val Glu Asp Ile Pro Arg Val 
        195                 200                 205             
Ile Gln Glu Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val 
    210                 215                 220                 
Leu Val Asp Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val 
225                 230                 235                 240 
Trp Asp Thr Ser Met Asn Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys 
                245                 250                 255     
Pro Pro Ala Thr Glu Leu Leu Glu Gln Val Leu Arg Leu Val Gly Glu 
            260                 265                 270         
Ser Arg Arg Pro Ile Leu Tyr Val Gly Gly Gly Cys Ser Ala Ser Gly 
        275                 280                 285             
Asp Glu Leu Arg Trp Phe Val Glu Leu Thr Gly Ile Pro Val Thr Thr 
    290                 295                 300                 
Thr Leu Met Gly Leu Gly Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu 
305                 310                 315                 320 
Arg Met Leu Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp 
                325                 330                 335     
Lys Ala Asp Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val 
            340                 345                 350         
Thr Gly Lys Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile 
        355                 360                 365             
Asp Ile Asp Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His Val Ser 
    370                 375                 380                 
Ile Cys Ala Asp Val Lys Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu 
385                 390                 395                 400 
Gln Gln Ser Thr Thr Lys Thr Ser Ser Asp Phe Ser Ala Trp His Asn 
                405                 410                 415     
Glu Leu Asp Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe 
            420                 425                 430         
Gly Glu Glu Ile Pro Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu 
        435                 440                 445             
Thr Lys Gly Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met 
    450                 455                 460                 
Trp Ala Ala Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser 
465                 470                 475                 480 
Ser Ala Gly Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala Gly 
                485                 490                 495     
Ala Ser Val Ala Asn Pro Gly Val Thr Val Val Asp Ile Asp Gly Asp 
            500                 505                 510         
Gly Ser Phe Leu Met Asn Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu 
        515                 520                 525             
Asn Leu Pro Val Lys Val Met Val Leu Asn Asn Gln His Leu Gly Met 
    530                 535                 540                 
Val Val Gln Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr 
545                 550                 555                 560 
Tyr Leu Gly Asn Pro Glu Cys Glu Ser Glu Ile Tyr Pro Asp Phe Val 
                565                 570                 575     
Thr Ile Ala Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys 
            580                 585                 590         
Ser Glu Val Arg Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro 
        595                 600                 605             
Tyr Leu Leu Asp Ile Ile Val Pro His Gln Glu His Val Leu Pro Met 
    610                 615                 620                 
Ile Pro Ser Gly Gly Ala Phe Lys Asp Met Ile Leu Asp Gly Asp Gly 
625                 630                 635                 640 
Arg Thr Val Tyr 
                

<210>  5
<211>  20
<212>  DNA
<213>  人工序列(Artificial Sequence)

<400>  5
tccttgaatg cgcccccact                                                    20

<210>  6
<211>  19
<212>  DNA
<213>   ALS-1F(Artificial Sequence)

<400>  6
atccgcattg agaacctcc                                                     19

<210>  7
<211>  20
<212>  DNA
<213>   ALS-4R(Artificial Sequence)

<400>  7
atgtccttga atgcgccacc                                                    20

<210>  9
<211>  20
<212>  DNA
<213>   ALS-6R(Artificial Sequence)

<400>  9
atgtccttga atgcgcctca                                                    20

<210>  10
<211>  20
<212>  DNA
<213>  ALS5-F(Artificial Sequence)

<400>  10
tcgcccaaac ccagaaaccc                                                    20

<210>  11
<211>  22
<212>  DNA
<213>  ALS5-R(Artificial Sequence)

<400>  11
ctctttatgg gtcattcagg tc                                                 22

<210>  12
<211>  24
<212>  DNA
<213>  ALS-U3-F(Artificial Sequence)

<400>  12
ggcatccttg aatgcgcccc cact                                               24

<210>  13
<211>  24
<212>  DNA
<213>  ALS-U3-R(Artificial Sequence)

<400>  13
aaacagtggg ggcgcattca agga                                               24

<210>  14
<211>  22
<212>  DNA
<213>  U-F(Artificial Sequence)

<400>  14
ctccgtttta cctgtggaat cg                                                 22

<210>  15
<211>  20
<212>  DNA
<213>  gRNA-R(Artificial Sequence)

<400>  15
cggaggaaaa ttccatccac                                                    20

<210>  16
<211>  38
<212>  DNA
<213>  Uctcg-B1(Artificial Sequence)

<400>  16
ttcagaggtc tctctcgcac tggaatcggc agcaaagg                                38

<210>  17
<211>  37
<212>  DNA
<213>  gRcggt-BL(Artificial Sequence)

<400>  17
agcgtgggtc tcgaccgggt ccatccactc caagctc                                 37

<210>  18
<211>  20
<212>  DNA
<213>  ALST-F(Artificial Sequence)

<400>  18
cgcatacata cttgggcaac                                                    20

<210>  19
<211>  22
<212>  DNA
<213>  ALST-R(Artificial Sequence)

<400>  19
acaaacatca taggcatacc ac                                                 22

<210>  20
<211>  20
<212>  DNA
<213>  ALS-F(Artificial Sequence)

<400>  20
tcgcccaaac ccagaaaccc                                                    20

<210>  21
<211>  22
<212>  DNA
<213>  ALS-R(Artificial Sequence)

<400>  21
ctctttatgg gtcattcagg tc                                                 22

<210>  22
<211>  19
<212>  DNA
<213>  hyg283-F(Artificial Sequence)

<400>  22
tccggaagtg cttgacatt                                                     19

<210>  23
<211>  19
<212>  DNA
<213>  hyg283-R(Artificial Sequence)

<400>  23
gtcgtccatc acagtttgc                                                     19

<210>  24
<211>  22
<212>  DNA
<213>  Cas9T-F(Artificial Sequence)

<400>  24
agcggcaaga ctatcctcga ct                                                 22

<210>  25
<211>  22
<212>  DNA
<213>  Cas9T-R(Artificial Sequence)

<400>  25
tcaatcctct tcatgcgctc cc                                                 22

<210>  27
<211>  21
<212>  DNA
<213>  ALS-1R(Artificial Sequence)

<400>  27
taggattacc atgccaagca c                                                  21

<210>  29
<211>  20
<212>  DNA
<213>  ALS-2R(Artificial Sequence)

<400>  29
atgtccttga atgcgccccc                                                    20

<210>  28
<211>  20
<212>  DNA
<213>  ALS-3R(Artificial Sequence)

<400>  28
atgtccttga atgcgcctcc                                                    20

<210>  28
<211>  20
<212>  DNA
<213>  ALS-4R(Artificial Sequence)

<400>  28
atgtccttga atgcgccacc                                                    20

<210>  29
<211>  20
<212>  DNA
<213>  ALS-5R(Artificial Sequence)

<400>  29
atgtccttga atgcgcccca                                                    20

<210>  30
<211>  20
<212>  DNA
<213>  ALS-6R(Artificial Sequence)

<400>  30
atgtccttga atgcgcctca                                                    20

<210>  31
<211>  20
<212>  DNA
<213>  ALS-7R(Artificial Sequence)

<400>  31
atgtccttga atgcgccaca                                                    20
     
