                         SEQUENCE LISTING

<110>  NATIONAL INSTITUTE OF PLANT GENOME RESEARCH
       NATIONAL INSTITUTE OF PLANT GENOME RESEARCH
 
<120>  POLYNUCLEOTIDE ASSOCIATED WITH ERGOSTEROL BIOSYNTHESIS AND USES 
       THEREOF

<130>  PD014201PCT

<160>  16    

<170>  PatentIn version 3.5

<210>  1
<211>  816
<212>  DNA
<213>  Artificial sequence

<220>
<223>  cDNA fragment encoding delta-7-sterol-C-5-desaturase activity 
       from tomato.

<400>  1
atggacgact acttgaattt atttgtagag gagacgtcgt tttttaatag gattgttctg       60

ggcacattct tgccggaatc atggtgggga ccactccctc attggtttca aggatggctt      120

cgcaactaca tcggtggagt cttgctctac tttatctctg ggtttctttg gtgcttctac      180

atctactact taaaacgcaa tgtttatcta cctaaagatg ccatcccttc aaatagagct      240

atgctcttgc aaataggagt tgcaatgaaa gctatgccat ggtactgtgc ccttccatca      300

ctttctgagt acatgattga aaatgggtgg actaaatgtt tctcaagaat aagcgatgtc      360

ggatggatac cctaccttat ctatgttgca gtttatctgg taatagtaga atttggtatc      420

tattggatgc atcgagagtt gcatgacata aaacctctgt acaaatatct ccatgctaca      480

catcatatat acaacaagca aaatacactc tccccatttg ctggtttggc attccaccca      540

ttggatggaa tactacaggc agtgccacac gtaatagctc tttttctgtt gcctgtgcat      600

ttcaccacac acattgctct cttattcata gaagccatat ggactgcaaa tattcatgac      660

tgtatacatg cgaaggtttg gcctgtaatg ggtgccggtt accataccat ccaccatact      720

acgtaccgcc ataattatgg tcactacaca atatggatgg actggatgtt tggaactctt      780

cgcgatcctg ttgaagatga agtgaagaaa ctgtaa                                816


<210>  2
<211>  271
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Amino acid sequence of a polypeptide having delta 
       7-sterol-C-5-desaturase activity encoded by a cDNA fragment 
       having nucleotide sequence as set forth in SEQ ID NO: 1

<400>  2

Met Asp Asp Tyr Leu Asn Leu Phe Val Glu Glu Thr Ser Phe Phe Asn 
1               5                   10                  15      


Arg Ile Val Leu Gly Thr Phe Leu Pro Glu Ser Trp Trp Gly Pro Leu 
            20                  25                  30          


Pro His Trp Phe Gln Gly Trp Leu Arg Asn Tyr Ile Gly Gly Val Leu 
        35                  40                  45              


Leu Tyr Phe Ile Ser Gly Phe Leu Trp Cys Phe Tyr Ile Tyr Tyr Leu 
    50                  55                  60                  


Lys Arg Asn Val Tyr Leu Pro Lys Asp Ala Ile Pro Ser Asn Arg Ala 
65                  70                  75                  80  


Met Leu Leu Gln Ile Gly Val Ala Met Lys Ala Met Pro Trp Tyr Cys 
                85                  90                  95      


Ala Leu Pro Ser Leu Ser Glu Tyr Met Ile Glu Asn Gly Trp Thr Lys 
            100                 105                 110         


Cys Phe Ser Arg Ile Ser Asp Val Gly Trp Ile Pro Tyr Leu Ile Tyr 
        115                 120                 125             


Val Ala Val Tyr Leu Val Ile Val Glu Phe Gly Ile Tyr Trp Met His 
    130                 135                 140                 


Arg Glu Leu His Asp Ile Lys Pro Leu Tyr Lys Tyr Leu His Ala Thr 
145                 150                 155                 160 


His His Ile Tyr Asn Lys Gln Asn Thr Leu Ser Pro Phe Ala Gly Leu 
                165                 170                 175     


Ala Phe His Pro Leu Asp Gly Ile Leu Gln Ala Val Pro His Val Ile 
            180                 185                 190         


Ala Leu Phe Leu Leu Pro Val His Phe Thr Thr His Ile Ala Leu Leu 
        195                 200                 205             


Phe Ile Glu Ala Ile Trp Thr Ala Asn Ile His Asp Cys Ile His Ala 
    210                 215                 220                 


Lys Val Trp Pro Val Met Gly Ala Gly Tyr His Thr Ile His His Thr 
225                 230                 235                 240 


Thr Tyr Arg His Asn Tyr Gly His Tyr Thr Ile Trp Met Asp Trp Met 
                245                 250                 255     


Phe Gly Thr Leu Arg Asp Pro Val Glu Asp Glu Val Lys Lys Leu 
            260                 265                 270     


<210>  3
<211>  36
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone LeC5SD in to pGEMT vector.

<400>  3
gctctagaat ggacgactac ttgaatttat ttgtag                                 36


<210>  4
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone LeC5SD in to pGEMT vector.

<400>  4
cagagctctt acagtttctt cacttcatct tcaac                                  35


<210>  5
<211>  30
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone LeC5SD for construct 1 in to 
       pHANNIBAL vector.

<400>  5
ctcgagcaaa gatgatagac aaatgagccc                                        30


<210>  6
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone LeC5SD for construct 1 in to 
       pHANNIBAL vector.

<400>  6
gaattccttg ttgtatatat gatgtgtagc atgg                                   34


<210>  7
<211>  30
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone antisense fragment of LeC5SD for 
       construct 1 in to pHANNIBAL vector.

<400>  7
tctagacaaa gatgatagac aaatgagccc                                        30


<210>  8
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone antisense fragment of LeC5SD for 
       construct 1 in to pHANNIBAL vector.

<400>  8
ggatcccttg ttgtatatat gatgtgtagc atgg                                   34


<210>  9
<211>  30
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone fragment of LeC5SD for construct 2 
       in to pHANNIBAL vector.

<400>  9
ctcgagcaac aagcaaaata cactctcccc                                        30


<210>  10
<211>  28
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone fragment of LeC5SD for construct 2 
       in to pHANNIBAL vector.

<400>  10
gaattcgcat caagagcatt tcacagta                                          28


<210>  11
<211>  30
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone antisense fragment of LeC5SD for 
       construct 2 in to pHANNIBAL vector.

<400>  11
tctagacaac aagcaaaata cactctcccc                                        30


<210>  12
<211>  28
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone antisense fragment of LeC5SD for 
       construct 2 in to pHANNIBAL vector.

<400>  12
ggatccgcat caagagcatt tcacagta                                          28


<210>  13
<211>  36
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Forward primer used to clone LeC5SD gene into pBI121 vector.

<400>  13
gctctagaat ggacgactac ttgaatttat ttgtag                                 36


<210>  14
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Reverse primer used to clone LeC5SD gene into pBI121 vector.

<400>  14
cagagctctt acagtttctt cacttcatct tcaac                                  35


<210>  15
<211>  1559
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Sequence of hairpin structure generated by construct 1.

<400>  15
ctcgagcagc tacacagcca gatcacaaca attaacacta ctagttgtga caagtcaaca       60

aagaaatcta aaaaattgtg tgccccacta acctctttgt tactttctcc aaatctctct      120

cttctccgat catctcccat cggaatctac tgaatcgtcg atggacgact acttgaattt      180

atttgtagag gagacgtcgt tttttaatag gattgttctg ggcacattct tgccggaatc      240

atggtgggga ccactccctc attggtttca aggatggctt cgcaactaca tcggtggagt      300

cttgctctac tttatctctg ggtttctttg gtgcttctac atctactact taaaacgcaa      360

tgtttatcta cctaaaggaa ttcccaattg gtaaggaaat aattattttc ttttttcctt      420

ttagtataaa atagttaagt gatgttaatt agtatgatta taataatata gttgttataa      480

ttgtgaaaaa ataatttata aatatattgt ttacataaac aacatagtaa tgtaaaaaaa      540

tatgacaagt gatgtgtaag acgaagaaga taaaagttga gagtaagtat attattttta      600

atgaatttga tcgaacatgt aagatgatat actagcatta atatttgttt taatcataat      660

agtaattcta gctggtttga tgaattaaat atcaatgata aaatactata gtaaaaataa      720

gaataaataa attaaaataa tattttttta tgattaatag tttattatat aattaaatat      780

ctataccatt actaaatatt ttagtttaaa agttaataaa tattttgtta gaaattccaa      840

tctgcttgta atttatcaat aaacaaaata ttaaataaca agctaaagta acaaataata      900

tcaaactaat agaaacagta atctaatgta acaaaacata atctaatgct aatataacaa      960

agcgcaagat ctatcatttt atatagtatt attttcaatc aacattctta ttaatttcta     1020

aataatactt gtagttttat taacttctaa atggattgac tattaattaa atgaattagt     1080

cgaacatgaa taaacaaggt aacatgatag atcatgtcat tgtgttatca ttgatcttac     1140

atttggattg attacagttg ggaaattggg ttcgaatcta gactttaggt agataaacat     1200

tgcgttttaa gtagtagatg tagaagcacc aaagaaaccc agagataaag tagagcaaga     1260

ctccaccgat gtagttgcga agccatcctt gaaaccaatg agggagtggt ccccaccatg     1320

attccggcaa gaatgtgccc agaacaatcc tattaaaaaa cgacgtctcc tctacaaata     1380

aattcaagta gtcgtccatc gacgattcag tagattccga tgggagatga tcggagaaga     1440

gagagatttg gagaaagtaa caaagaggtt agtggggcac acaatttttt agatttcttt     1500

gttgacttgt cacaactagt agtgttaatt gttgtgatct ggctgtgtag ctgggatcc      1559


<210>  16
<211>  1661
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Sequence of hairpin structure generated by construct 2.

<400>  16
ctcgagtgct ctcttattca tagaagccat atggactgca aatattcatg actgtataca       60

tgcgaaggtt tggcctgtaa tgggtgccgg ttaccatacc atccaccata ctacgtaccg      120

ccataattat ggtcactaca caatatggat ggactggatg tttggaactc ttcgcgatcc      180

tgttgaagat gaagtgaaga aactgtaaca ttcatccctg taacgaattg ttattcatcg      240

tcttatttat taggttgcag agtggttggt gcttcattta gagtttgttg tgtgtgcatt      300

tcatggatgt aaattttact agtgggtttt caatgtttag acaaaaaata ttatccagga      360

aaaatactgt gaaatgctct tgatgcattt gaggtattgt tattgaggaa agaagcaaga      420

gatccaagga attcccaatt ggtaaggaaa taattatttt cttttttcct tttagtataa      480

aatagttaag tgatgttaat tagtatgatt ataataatat agttgttata attgtgaaaa      540

aataatttat aaatatattg tttacataaa caacatagta atgtaaaaaa atatgacaag      600

tgatgtgtaa gacgaagaag ataaaagttg agagtaagta tattattttt aatgaatttg      660

atcgaacatg taagatgata tactagcatt aatatttgtt ttaatcataa tagtaattct      720

agctggtttg atgaattaaa tatcaatgat aaaatactat agtaaaaata agaataaata      780

aattaaaata atattttttt atgattaata gtttattata taattaaata tctataccat      840

tactaaatat tttagtttaa aagttaataa atattttgtt agaaattcca atctgcttgt      900

aatttatcaa taaacaaaat attaaataac aagctaaagt aacaaataat atcaaactaa      960

tagaaacagt aatctaatgt aacaaaacat aatctaatgc taatataaca aagcgcaaga     1020

tctatcattt tatatagtat tattttcaat caacattctt attaatttct aaataatact     1080

tgtagtttta ttaacttcta aatggattga ctattaatta aatgaattag tcgaacatga     1140

ataaacaagg taacatgata gatcatgtca ttgtgttatc attgatctta catttggatt     1200

gattacagtt gggaaattgg gttcgaatct agacttggat ctcttgcttc tttcctcaat     1260

aacaatacct caaatgcatc aagagcattt cacagtattt ttcctggata atattttttg     1320

tctaaacatt gaaaacccac tagtaaaatt tacatccatg aaatgcacac acaacaaact     1380

ctaaatgaag caccaaccac tctgcaacct aataaataag acgatgaata acaattcgtt     1440

acagggatga atgttacagt ttcttcactt catcttcaac aggatcgcga agagttccaa     1500

acatccagtc catccatatt gtgtagtgac cataattatg gcggtacgta gtatggtgga     1560

tggtatggta accggcaccc attacaggcc aaaccttcgc atgtatacag tcatgaatat     1620

ttgcagtcca tatggcttct atgaataaga gagcaggatc c                         1661


