添付資料1  ラウンドアップ耐性大豆遺伝子のリードスルーとスプライシングで生じたm−RNA(文献4より)

 

       1359

gDNA   TCGTGTCGGAAAACCCTGTCACGGTGGACGATGCCACGATGATCGCCACGAGCTTCCCGGAGTTCATGGACCTGATGGCCGGGCTGGGCGCGAAGATCGAACTCTCCGA
cDNA #3   TCGTGTCGGAAAACCCTGTCACGGTGGACGATGCCACGATGATCGCCACGAGCTTCCCGGAGTTCATG-----------------------------------------
cDNA #4   TCGTGTCGGAAAACCCTGTCACGGTGGACGATGCCACGATGATCGCCACGAGCTTCCCGGAGTTCATGGACCTGATGGCCGGGCTGGGCGCGAAGATCGAACTCTCCGA
cDNA #5   TCGTGTCGGAAAACCCTGTCACGGTGGACGATGCCACGATGATCGCCACGAGCTTCCCGGAGTTCATGGACCT------------------------------------
cDNA #6   TCGTGTCGGAAAACCCTGTCACGGTGGACGATGCCACGATGATCGCCACGAGCTTCCCGGAGTTCATGGACCT------------------------------------
 
          1468
gDNA      TACGAAGGCTGCCTGATGAGCTCGAATTCGAGCTCGGTACCGGATCCAATTCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTC
cDNA #3   -------------------------------------------------------------------------------------------------------------
cDNA #4   TACG---------------------------------------------------------------------------------------------------------
cDNA #5   -------------------------------------------------------------------------------------------------------------
cDNA #6   -------------------------------------------------------------------------------------------------------------
 
          1577
gDNA      TTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCA
cDNA #3   -------------------------------------------------------------------------------------------------------------
cDNA #4   -------------------------------------------------------------------------------------------------------------
cDNA #5   -------------------------------------------------------------------------------------------------------------
cDNA #6   -------------------------------------------------------------------------------------------------------------
           
          1686
gDNA      ATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGGATCGATCCCCCACCGG
cDNA #3   -------------------------------------------------------------------------------------------------------------
cDNA #4   -------------------------------------------------------------------------------------------------------------
cDNA #5   -------------------------------------------------------------------------------------------------------------
cDNA #6   -------------------------------------------------------------------------------------------------------------
 
          1795                                                                                                     1901
gDNA      TCCTTCATGTTCGGCGGTCTCGCGAGCGGTGAAACGCGCATCATTGGCCTTCTGGAAGGCGAGGACGTCATCAATACGGGCAAGGCCATGCAGGCCATGGGCGCCA
cDNA #3   ---------TTCGGCGGTCTCGCGAGCGGTGAAACGCGCATCATTGGCCTTCTGGAAGGCGAGGACGTCATCAATACGGGCAAGGCCATGCAGGCCATGGGCGCCA
cDNA #4   ------------------------------------------------------------------------------GGCAAGGCCATGCAGGCCATGGGCGCCA
cDNA #5   -----CATGTTCGGCGGTCTCGCGAGCGGTGAAACGCGCATCATTGGCCTTCTGGAAGGCGAGGACGTCATCAATACGGGCAAGGCCATGCAGGCCATGGGCGCCA
cDNA #6   --------------------------------------------------TCTGGAAGGCGAGGACGTCATCAATACGGGCAAGGCCATGCAGGCCATGGGCGCCA
 


注:

g DNA 挿入されたRR大豆DNA

     (1359から1901まで表示)

TGA(ボールド):CP4EPSPS ORFの停止コドン(1480

NOS配列(イタリック):15311831)

アンダーライン:PCR分析用プライマー結合部位 

36:スプライスで出来た新mRNA

の転写部分(---------が欠損部分)

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

添付資料2 CP4EPSPSとリードスルーで出来る融合たんぱく質のアミノ酸配列
 
                  1                                                                                                               100

CP4EPSPS           MLHGASSRPATARKSSGLSQTVRIPGDKSISHRSFMFGGLASGETRITGLLEGEDVINTGKAMGAMGARIRKEGDTWIIDGVGNGGLLAPEAPLDFGNAA

                   101                                                                                                           200

CP4EPSPS           TGCRLTMGLVGVYDFDSTFIGDASLTKRPMGRVLNPLREMGVGVKSEDGDRLPVTLRGPKTPTPITYRVPMASAGVKSAVLLAGLNTPGITTVIEPIMTR

     201                                                                                                           300

CP4EPSPS           DHTEKMLGGFGANLTVETDADGVRTIRLEGRGKLTGGVIDVPGDPSSTAFPLVAALLVPGSDVTILNVLMNPTRTGLILTLGEMGADIEVINPRLAGGED

                   301                                                                                                           400

CP4EPSPS           VADLRVRSSTLKGVTVPEDRAPSMIDEYPILAVAAAFAEGATVMNGLEELRVKESDRLSAVANGLKLNGVDCDEGETSLVVRGRPDGKGLGNASGAAVAT

      401                                                         455                                                                                 

CP4EPSPS           HLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFMDLMAGLGAKIELSDTKAA     1                                       40

CP4EPSPS #3        HLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFM-----------------------FGGLASGETRITGLLEGEDVINTGKAMGAMGARIRKEGDT

CP4EPSPS #4        HLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFMDLMAGLGAKIELSDT-------------------------------GKAMGAMGARIRKEGDT

CP4EPSPS #5        HLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFMDLM--------------------FGGLASGETRITGLLEGEDVINTGKAMGAMGARIRKEGDT

CP4EPSPS #6        HLDHRIAMSFLVMGLVSENPVTVDDATMIATSFPEFMDL-----------------------------------LEGEDVINTGKAMGAMGARIRKEGDT

 

                   41                                                                   100

CP4EPSPS #3        WIIDGVGNGGLLAPEAPLDFGNAATGCRLTMGLVGVYDFKRIMLGNFSEIISIFLGISAVTGE

CP4EPSPS #4        WIIDGVGNGGLLAPEAPLDFGNAATGCRLTMGLVGVYDFKRIMLGNFSEIISIFLGISAVTGE

CP4EPSPS #5        WIIDGVGNGGLLAPEAPLDFGNAATGCRLTMGLVGVYDFKRIMLGNFSEIISIFLGISAVTGE

CP4EPSPS #6        WIIDGVGNGGLLAPEAPLDFGNAATGCRLTMGLVGVYDFKRIMLGNFSEIISIFLGISAVTGE

 
 
 
説明:CP4EPSPS #3〜#6が新たな融合たんぱく質。401番目から始まり長さの異なる4種類の新規たんぱく質が可能
   アンダーライン部分は、融合たんぱく質内に取り込まれた、CP4EPSPSの一部の配列
   ボールドタイプの部分は宿主大豆由来の配列換えしたボーダーDNAによるアミノ酸配列
 
 
 
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