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):1531−1831)
アンダーライン:PCR分析用プライマー結合部位
#3〜6: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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