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<⾗ᢱ㧞> 㒰⨲ࠣࡎࠨ࠻⠴ᕈ߮ࠦ࠙࠴ࡘ࠙⋡߮࠴࡚࠙⋡ኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ (cp4 epsps, cry3Bb1, cry1Ab, Zea mays subsp. mays (L.) Iltis.) (MON 88017MON 810, OECD UI: MON-88Ø17-3MON-ØØ81Ø-6) ↳⺧ᦠ╬ߩⷐ ╙৻⒳↪ⷙ⒟ᛚ↳⺧ᦠ································································································· 1 ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩⷐ ╙৻ ↢‛ᄙ᭽ᕈᓇ㗀ߩ⹏ଔߦᒰߚࠅ㓸ߒߚᖱႎ ....................................................2 1 ኋਥߪኋਥߩዻߔࠆಽ㘃ቇߩ⒳ߦ㑐ߔࠆᖱႎ··········································· 2 (1) ಽ㘃ቇߩ⟎ઃߌ߮⥄ὼⅣႺߦ߅ߌࠆಽᏓ⁁ᴫ······································· 2 (2) ↪╬ߩᱧผ߮⁁·························································································· 2 (3) ↢ℂ⊛߮↢ᘒቇ⊛․ᕈ······················································································ 3 2 ㆮવሶ⚵឵߃↢‛╬ߩ⺞╬ߦ㑐ߔࠆᖱႎ ······················································ 5 (1) ଏਈᩭ㉄ߦ㑐ߔࠆᖱႎ·························································································· 5 (2) ࡌࠢ࠲ߦ㑐ߔࠆᖱႎ························································································ 17 (3) ㆮવሶ⚵឵߃↢‛╬ߩ⺞ᣇᴺ ········································································ 17 (4) ⚦⢩ౝߦ⒖ߒߚᩭ㉄ߩሽ⁁ᘒ߮ᒰᩭ㉄ߦࠃࠆᒻ (5) (6) 3 (1) (2) (3) (4) ⾰⊒ߩቯᕈ···································································································· 21 ㆮવሶ⚵឵߃↢‛╬ߩᬌ߮⼂ߩᣇᴺਗ߮ߦߘࠇࠄ ߩᗵᐲ߮ା㗬ᕈ································································································ 24 ኋਥߪኋਥߩዻߔࠆಽ㘃ቇߩ⒳ߣߩ⋧㆑················································· 24 ㆮવሶ⚵឵߃↢‛╬ߩ↪╬ߦ㑐ߔࠆᖱႎ ···················································· 32 ↪╬ߩౝኈ ······································································································· 32 ↪╬ߩᣇᴺ ······································································································· 32 ᛚࠍฃߌࠃ߁ߣߔࠆ⠪ߦࠃࠆ╙৻⒳↪╬ߩ㐿ᆎᓟߦ ߅ߌࠆᖱႎ㓸ߩᣇᴺ························································································ 32 ↢‛ᄙ᭽ᕈᓇ㗀߇↢ߕࠆ߅ߘࠇߩࠆ႐วߦ߅ߌࠆ↢‛ ᄙ᭽ᕈᓇ㗀ࠍ㒐ᱛߔࠆߚߩភ⟎ ···································································· 32 ታ㛎ቶ╬ߢߩ↪ߪ╙৻⒳↪╬߇੍ቯߐࠇߡࠆⅣ Ⴚߣ㘃ૃߩⅣႺߢߩ↪╬ߩ⚿ᨐ ···································································· 32 (6) ࿖ᄖߦ߅ߌࠆ↪╬ߦ㑐ߔࠆᖱႎ ···································································· 32 ╙ੑ 㗄⋡ߏߣߩ↢‛ᄙ᭽ᕈᓇ㗀ߩ⹏ଔ .................................................................... 34 1 ┹วߦ߅ߌࠆఝᕈ···························································································· 34 2 ኂ‛⾰ߩ↥↢ᕈ································································································ 36 3 㔀ᕈ ··················································································································· 46 4 ߘߩઁ ··················································································································· 47 ╙ਃ ↢‛ᄙ᭽ᕈᓇ㗀ߩ✚ว⊛⹏ଔ ............................................................................ 48 ᒁ↪ᢥ₂ ······························································································································ 50 ✕ᕆភ⟎⸘↹ᦠ ·················································································································· 51 (5) ╙ ৻ ⒳ ↪ ⷙ ⒟ ᛚ ↳ ⺧ ᦠ ᐔᚑ 16 ᐕ 10 19 ᣣ ㄘ ᨋ ᳓ ↥ ᄢ ⤿ ፉ ቱ િ Ლ Ⅳ Ⴚ ᄢ ⤿ ዊ ᳰ ⊖ ว ሶ Ლ ↳⺧⠪ ᳁ฬ ᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ ઍข✦ᓎ␠㐳 ጊᩮ ♖৻㇢ ශ ᚲ ᧲੩ㇺਛᄩ㌁ᐳ྾ৼ⋡ 10 ⇟ 10 ภ ╙৻⒳↪ⷙ⒟ߦߟߡᛚࠍฃߌߚߩߢޔㆮવሶ⚵឵߃↢‛╬ߩ↪╬ߩⷙߦ ࠃࠆ↢‛ߩᄙ᭽ᕈߩ⏕ߦ㑐ߔࠆᴺᓞ╙㧠᧦╙㧞㗄ߩⷙቯߦࠃࠅޔᰴߩߣ߅ࠅ↳⺧ߒ߹ ߔޕ ㆮવሶ⚵឵߃↢‛╬ߩ 㒰⨲ࠣࡎࠨ࠻⠴ᕈ߮ࠦ࠙࠴ࡘ࠙⋡߮࠴࡚࠙⋡ኂ ⒳㘃ߩฬ⒓ ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ(cp4 epsps, cry3Bb1, cry1Ab, Zea mays subsp. mays (L.) Iltis)(MON 88017 MON 810, OECD UI: MON-88Ø17-3MON-ØØ81Ø-6) ㆮવሶ⚵឵߃↢‛╬ߩ 㘩↪ߪ㘺ᢱ↪ߦଏߔࠆߚߩ↪ޔᩱၭޔടᎿޔ▤ޔ ╙৻⒳↪╬ߩౝኈ ㆇ៝߮ᑄ᫈ਗ߮ߦߎࠇࠄߦઃ㓐ߔࠆⴕὑ ㆮવሶ⚵឵߃↢‛╬ߩ 㧙 ╙৻⒳↪╬ߩᣇᴺ 1 ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩⷐ ╙৻↢‛ᄙ᭽ᕈᓇ㗀ߩ⹏ଔߦᒰߚࠅ㓸ߒߚᖱႎ 1 ኋਥߪኋਥߩዻߔࠆಽ㘃ቇߩ⒳ߦ㑐ߔࠆᖱႎ (1) ಽ㘃ቇߩ⟎ઃߌ߮⥄ὼⅣႺߦ߅ߌࠆಽᏓ⁁ᴫ ࠗ㧚৻⥸ߦ࠻࠙ࡕࡠࠦࠪߩቇฬߪ Zea mays L.ߢࠆ߇ޔㄭᐕߩࠪࠦࡠࡕ࠙࠻ޔ ㄭ✼⒳ߢࠆ৻ᐕ↢࠹ࠝࠪࡦ࠻߇ Z. mays ߦಽ㘃ߐࠇߚ⚿ᨐߩߘߪࠪࠦࡠࡕ࠙࠻ޔ ⒳ߣߒߡ Z. mays subsp. mays (L.) Iltis ߣߒߡಽ㘃ߐࠇࠆࠃ߁ߦߥߞߚޕ ࡠ㧚ኋਥߪࠗࡀ⑼(Gramineae)࠻࠙ࡕࡠࠦࠪዻ(Zea)ߦዻߔࠆ࠻࠙ࡕࡠࠦࠪ(Zea mays)ߢ࠻ࡦ࠺ޔ⒳ߦዻߔࠆޕ ࡂ㧚ේ↥ߦߟߡߪ☨߷߶ޔ࿖ߩධㇱޔࠦࠪࠠࡔޔਛ☨ࠆߪධ☨ߦ߆ ߌߡߩၞߣ⠨߃ࠄࠇࠆ߇ޔቯ⊛ߥ⺑ߪߥߊⶄࠄࠇߎޔᢙၞ߇ߘࠇߙࠇ⁛┙ ߒߚḮߢࠆߣߔࠆ⺑ߣࠦࠪࠠࡔޔධㇱන⁛ࠍේߣߔࠆ⺑߇ࠆ(ᢥ₂ 1)ޕዏޔ ࠊ߇࿖ߦ߅ߌࠆ⥄ὼಽᏓߩႎ๔ߪߥޕ (2) ↪╬ߩᱧผ߮⁁ ࠗ㧚࠻࠙ࡕࡠࠦࠪߩᦨฎߩᩱၭḮߪ߆ࠄ 9,000 ᐕ೨ߣߐࠇߡࠆ(ᢥ₂ 1)ޕ ߘߩᓟޔේ᳃ߩᚻߦࠃࠅ⢒⒳ޔຠ⒳ᡷ⦟߇ⴕࠊࠇ♿ޔర೨ 3000 ᐕ㨪1500 ᐕ㗃 ߦߪޔઍߩᩱၭဳߦㄭ࠻࠙ࡕࡠࠦࠪ߇ᧄᩰ⊛ߦᩱၭߐࠇࠆࠃ߁ߦߥࠅޔධർ ࠕࡔࠞᄢ㒽ߩฦߦવߒߩߘޔવߩㆊ⒟ߢߐࠄߦ࠺ࡦ࠻ࠗࠬޔࡊ࠶ࡐޔ ࠻⒳ߥߤߩᄙᢙߩᄌ⇣⒳߇↢ߓߚߣ⠨߃ࠄࠇߡࠆ(ᢥ₂ 2)ޕᣣᧄ߳ߪᄤᱜᐕ㑆 (1579 ᐕ)ߦ㐳ፒ߆྾࿖ߦવ᧪ߒߚߩ߇ᦨೋߢࠆߣߐࠇޔᩱၭߩᱧผߪ㐳ޕ ࡠ㧚ޔ㘺ᢱߣߒߡߩ↪߇ਥᵹߢࠆ߇ޔ㘩↪ޔ㘩↪ᴤޔố☳ߥߤߩ㘩ຠ ߣߒߡߩ↪ㅜ߽ᄙጘߦࠊߚࠆ(ᢥ₂ 2; ᢥ₂ 1)ޕߪࠪࠦࡠࡕ࠙࠻ޔ⇇ߢᦨ߽ ᐢߊᩱၭߐࠇߡࠆⓃ‛ߢ☨ޔ࿖ޔਛ࿖ࡦ࠴ࡦ࡞ࠕޔ࡞ࠫࡉޔ࡛߮ࡠ࠶ ࡄ⻉࿖ߥߤࠍਛᔃߦޔർ✲ 58 ᐲ߆ࠄධ✲ 40 ᐲߦ⥋ࠆ▸࿐ߢᩱၭน⢻ߢࠆ(ᢥ₂ 3; ᢥ₂ 1)ޕ࿖ㅪ㘩♳ㄘᬺᯏ㑐(FAO)ߩ⛔⸘ᖱႎߦၮߠߊߣޔ2002 ᐕߦ߅ߌࠆో ⇇ߩ࠻࠙ࡕࡠࠦࠪߩᩱၭ㕙Ⓧߪ⚂ 1 ం 4 ජਁ ha ߢࠅޔ࿖ࠍߍࠆߣ☨࿖߇ 2,800 ਁ haޔਛ࿖߇ 2,500 ਁ ha ߇࡞ࠫࡉޔ1,200 ਁ ha ߇ࠦࠪࠠࡔޔ700 ਁ haޔ ࠗࡦ࠼߇ 600 ਁ ha ߇ࠕࠚࠫࠗ࠽ޔ400 ਁ haޔධࠕࡈࠞ߇ 300 ਁ ha ߣߥߞߡ ࠆ(ᢥ₂ 4)ޕዏޔห⛔⸘ᖱႎߦၮߠߊ 2002 ᐕߩᣣᧄߦ߅ߌࠆᩱၭ㕙Ⓧߪ⚂ 3 ਁ ha ߢߞߚޕ 2 ᣣᧄߪᶏᄖ߆ࠄ⚂ 1,600 ਁ࠻ࡦߩ࠻࠙ࡕࡠࠦࠪࠍ㘺ᢱ↪ޔ㘩ຠ↪ߣߒߡャߒ ߡࠆޕ㘺ᢱ↪ߪ⚂ 1,100 ਁ࠻ࡦޔ㘩ຠ↪ߪ⚂ 500 ਁ࠻ࡦߢਥߥ↪ㅜߪố☳⇣ޔ ᕈൻ♧ߢࠆޕ ࠊ߇࿖ߢߩ㘺ᢱ↪࠻࠙ࡕࡠࠦࠪߩᘠⴕᩱၭᴺߪએਅߩߣ߅ࠅߢࠆޕ⒳ㆡᦼ ߪኙ߆ࠄ᷷ᥦ߹ߢߪ 5 ৻ޔㇱߩᥦߢߪ 4 ߆ࠄ 6 ߹ߢߢࠆޕㆡᱜᩱ ᬀኒᐲߪ 10a ߚࠅ 6,000㨪8,000 ᧄߢࠆޕ㔀⨲㒐㒰ߩߚ↢ޔ⢒ೋᦼߦ㒰⨲ ᢔᏓ߿ 2㨪3 ࿁ߩਛ⠹ၭᬺࠍⴕ߁ޕ㓽Ⓞߩࠃࠅ 35㨪45 ᣣᓟߩ㤛ᾫᦼߦ ㇱࠍⓠߔࠆ(ᢥ₂ 3)ޕ ዏޔ࿖ౝਥⷐ⒳⧣ࡔࠞߩຠ⒳ࠬ࠻ߦၮߠߊߣޔߦ⥸৻ޔᩱၭ↪ߣߒ ߡᏒ⽼ߐࠇߡࠆ࠻࠙ࡕࡠࠦࠪߩ߶ߣࠎߤోߡߪ৻ઍ㔀⒳ຠ⒳(F1)ߥߩߢޔⓠ ⒳ሶ߇⠉ᐕߦᩱၭ↪ߣߒߡ⒳ߐࠇࠆߎߣߪ৻⥸⊛ߢߥޕ (3) ↢ℂ⊛߮↢ᘒቇ⊛․ᕈ ࠗ ၮᧄ⊛․ᕈ 㧙 ࡠ ↢ᕷߪ↢⢒น⢻ߥⅣႺߩ᧦ઙ ࠻࠙ࡕࡠࠦࠪ⒳ሶߩ⊒⧘ㆡ᷷ߪ 32㨪36͠ᦨޔૐ⊒⧘᷷ᐲ߮ᦨૐ↢⢒᷷ᐲߪ 6 㨪10͠ߢࠅޔታ㓙ߦߪ 13㨪14͠એߩᤨᦼ߇⒳ㆡᦼߣߐࠇޔຠ⒳߿ၞߦࠃ ߞߡᩱၭᤨᦼߪᄙዋ⇣ߥࠆ߇ޔਥߦᤐߦ⒳ߐࠇߡ⑺ߦⓠߐࠇࠆ৻ᐕ↢ߩ‛ ߢࠆ(ᢥ₂ 3)ߦ⥸৻ޔߚ߹ޕ⍴ᣣᬀ‛ߢࠅߩߘޔᗵశᕈߪ᥅↢⒳߶ߤᢅᗵߢޔ ᣧ↢ຠ⒳߶ߤ㊰ᗵߢࠆ(ᢥ₂ 3)ࠄࠇߎޕ᷷ᐲ᧦ઙ╬ߩઁ࠻ࡦ࠺ޔ⒳ߩ႐วߪ⒳ሶ ㊀㊂ߩ 70%ߩ᳓ࠍๆ߁ߣ⊒⧘ߔࠆ(ᢥ₂ 5)ߩࠪࠦࡠࡕ࠙࠻ޔߚ߹ޕᩱၭߦߪ⣣ᬀߦ ንფ߇ㆡߒޔpH5.5㨪8.0 ߩ▸࿐ߢᩱၭน⢻ߢࠆ(ᢥ₂ 5)ޕ ߩ࠻࠙ࡕࡠࠦࠪߪᩱၭ‛ߣߒߡ㜞ᐲߦੱὑ⊛ߦࠄࠇߚ‛ߢࠅ⥄ޔ ὼ᧦ઙਅߢᬀ‛ߣߒߡ❥ᱺߒ↢ޔሽߔࠆߚߩ⢻ജߪᄬࠊࠇߡࠆ(ᢥ₂ 6; ᢥ₂ 1)ޕ ࡂ 㘩ᕈߪነ↢ᕈ 㧙 3 ࠾ ❥ᱺߪჇᱺߩ᭽ᑼ Ԙቢᾫߒߚ⒳ሶߪ㓽Ⓞߩ൮⊹ߢⷒࠊࠇߡ߅ࠅ⥄ޔὼߩ⣕☸ᕈߪߥ(ᢥ₂ 1)࠻ޕ ࠙ࡕࡠࠦࠪߪ㐳㑆ᩱၭᬀ‛ൻߐࠇߡߚߚߦޔ㊁↢ߣߒߡ↢߈ᱷࠆ⢻ജࠍᄬ ߞߡ߅ࠅߩߘޔ⒳ሶࠍಽᢔߐߖࠆߚߦߪੱ㑆ߩખ߇ᔅⷐߢࠆޕ⒳ሶߩભ⌁ ᕈߪᭂߡૐߊޔⓠᤨߦ⒳ሶ߇ߦ⪭ਅߒߡ߽ޔფ᷷ᐲ߇ 10͠ߦ㆐ߔࠆ߹ ߢ⊒⧘ߒߥߚޔᄙߊߩ႐วࠆߔ⧘⊒ޔ೨ߦ⣣ᢌߒᨗᱫߔࠆ(ᢥ₂ 2; ᢥ₂ 3)ޕ ߹ߚޔߦ⊒⧘ߒߡ߽↢㐳ὐ߇ㇱߦࠆೋᦼ↢⢒ᤨ(5㨪7 ⪲ᦼ)ߦޔ0͠એਅߢ 6㨪8 ᤨ㑆એߩ᧦ઙਅߦ߅߆ࠇࠆߣ↢ሽߢ߈ߥ(ᢥ₂ 1)ޕ⒳ሶߩኼߪᏱ᷷ ሽߢߪ⍴ߊޔ2 ᐕ⋡߆ࠄ⊒⧘₸߇ૐਅߔࠆޕ ԙ࠻࠙ࡕࡠࠦࠪߪᩕ㙃❥ᱺߪߖߕޔ⒳ሶ❥ᱺߔࠆ⥄ޕὼ᧦ઙߦ߅ߡᬀ‛ࠍ ౣ↢ߒ߁ࠆ⚵❱ߪེቭ߆ࠄߩ⧘․ᕈ߇ࠆߣ߁ႎ๔ߪߎࠇ߹ߢߩߣߎࠈߥ ޕ Ԛ࠻࠙ࡕࡠࠦࠪߪ㓽㓶หᩣᬀ‛ߩ৻ᐕ↢‛ߢߥ⊛ဳౖޔ㘑ᇦ⧎ߢࠅߣ߶ޔ ࠎߤߪઁኅฃ☳ߦࠃߞߡࠄࠇߚ⒳ሶߦࠃࠅ❥ᱺߔࠆ߇⥄ޔኅਇวᕈ߇ߥߚ ⥄ኅฃ☳߽น⢻ߢࠆ(ᢥ₂ 1; ᢥ₂ 7)ߩࠪࠦࡠࡕ࠙࠻ޕㄭ✼⒳ߪ Tripsacum ዻ ߣ Zea ዻߦಽ㘃ߐࠇࠆ࠹ࠝࠪࡦ࠻ߢࠆ߇⥄ߣࠪࠦࡠࡕ࠙࠻ޔὼ㔀น⢻ߥߩߪ ࠹ࠝࠪࡦ࠻ߩߺߢޔTripsacum ዻߣߩ⥄ὼ㔀ߪ⍮ࠄࠇߡߥ(ᢥ₂ 1)ࠪࠝ࠹ޕ ࡦ࠻ߪࡔࠠࠪࠦߣࠣࠔ࠹ࡑߦߩߺ⥄ὼಽᏓߒߡ߅ࠅ৻ޔᣇޔTripsacum ዻߩಽᏓ ၞߪർࠕࡔࠞ᧲ධㇱ᧲ࠬ࠺ࡦࠕߩߡߌ߆ߦࠕࡆࡏࠄ߆ࠕࡆࡦࡠࠦޔߩૐ ߩߎޔߡߒߘޔዻߩਛᔃߣ⠨߃ࠄࠇࠆࡔࠠࠪࠦ ߩࡑ࠹ࠔࠣޔ3 ၞߦᄢ ߐࠇߡࠆ(ᢥ₂ 2; ᢥ₂ 3; ᢥ₂ 1; ᢥ₂ 8)ޕ ᚒ߇࿖ߢߪޔ ࠹ࠝࠪࡦ࠻߮ Tripsacum ዻߩ㊁↢⒳ߪႎ๔ߐࠇߡߥ(ᢥ₂ 9; ᢥ₂ 3)ޕ ԛ࠻࠙ࡕࡠࠦࠪߩ৻ᧄߩ㓶Ⓞߦߪ 1,200㨪2,000 ߩዊⓄ߇ࠅޔ 1,600 ਁ㨪3,000 ਁߩ⧎☳☸ࠍᒻᚑߔࠆߩ☳⧎ޕኼߪ⋓ᄐߩ߶႐᧦ઙਅߢߪ 24 ᤨ㑆એౝߢࠆ ߇ޔⅣႺߦࠃࠅ 2 ᤨ㑆߆ࠄ 8 ᣣ߹ߢߩ߇ࠆ(ᢥ₂ 10)ߪ☳⧎ޕᒻߢ⋥ޔᓘߪ 90-100Ǵm ߢࠆ(ᢥ₂ 11)ޕ 㘑ᇦߦࠃࠆઁኅฃ☳߇ਥߢࠆ߇᥉ㅢߩ߶႐ߢ 1㨪5% ߩ⥄ኅฃ☳߇߈ࠆޕ㓶Ⓞߩ㐿⧎ߦࠃߞߡ㘧ᢔߒߚ⧎☳ߪޔ㓽Ⓞ߆ࠄߒߚ⛚ ♻ߦઃ⌕ߒߡ⊒⧘ߒޔ24 ᤨ㑆એౝߦฃ♖ࠍቢੌߔࠆ(ᢥ₂ 1)ࠦࡠࡕ࠙࠻ޔߚ߹ޕ ࠪ⧎☳߇㘧ᢔߔࠆ〒㔌ߪޔᨋޔጊߥߤߩㆤ⭁‛ߩήޔ㘑ะ߈ߥߤߢ⇣ߥࠆ߇ޔ ߅ࠃߘ 300㨪500m ߣߐࠇߡࠆ(ᢥ₂ 3)ޕ ࡎ ∛ේᕈ 㧙 4 ࡋ ኂ‛⾰ߩ↥↢ᕈ ࠻࠙ࡕࡠࠦࠪߦ߅ߡ⥄ޔὼ᧦ઙਅߢ࿐ߩ㊁↢േᬀ‛╬ߩ↢⢒߹ߚߪ↢ᕷߦ ᓇ㗀ࠍ߷ߔኂ‛⾰ߩ↥↢ߪႎ๔ߐࠇߡߥޕ ࠻ ߘߩઁߩᖱႎ ߎࠇ߹ߢޔㆇ៝╬ߦ߅ߡߎ߷ࠇ⪭ߜߚ࠻࠙ࡕࡠࠦࠪ߇⇌એᄖߢ↢⢒ߒߚߣ ߁ႎ๔ߪߥޕ 2 ㆮવሶ⚵឵߃↢‛╬ߩ⺞╬ߦ㑐ߔࠆᖱႎ 㒰⨲ࠣࡎࠨ࠻߮ࠦ࠙࠴ࡘ࠙⋡ኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ (cp4 epsps, cry3Bb1, Zea mays subsp. mays (L.) Iltis) (MON 88017, OECD UI: MON-88Ø17-3)(એਅޔ MON 88017 ߣߔࠆ)ߣ࠴࡚࠙⋡ኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ (cry1Ab, Zea mays subsp. mays (L.) Iltis) (MON 810, OECD UI: MON-ØØ81Ø-6)(એਅޔMON 810 ߣߔࠆ)ࠍᓥ ᧪ߩ㔀⢒⒳ᴺࠍ↪ߡ㈩ߐߖߚ㒰⨲ࠣࡎࠨ࠻⠴ᕈ߮ࠦ࠙࠴ࡘ࠙⋡ ߮࠴࡚࠙⋡ኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ (cp4 epsps, cry3Bb1, cry1Ab, Zea mays subsp. mays (L.) Iltis) (OECD UI: MON-88Ø17-3MON-ØØ81Ø-6) (એਅ♽ࠢ࠶࠲ࠬᧄޟޔ ⛔࠻࠙ࡕࡠࠦࠪ ࠆߢ⛔♽ⷫޔߪ)ࠆߔߣޠMON 88017 ߣ MON 810 ߩ 2 ߟߩ⚵឵ ߃࠻࠙ࡕࡠࠦࠪߩߘࠇߙࠇߩ․ᕈࠍߔࠆޔߡߞ߇ߚߒޕએਅߢߪ MON 88017 ߣ MON 810 ߩ⺞╬ߦ㑐ߔࠆᖱႎߦߟߡߦㅀߴߚޕ (1) ଏਈᩭ㉄ߦ㑐ߔࠆᖱႎ ࠗ ᭴ᚑ߮᭴ᚑⷐ⚛ߩ↱᧪ MON 88017 ߩߦ↪ࠄࠇߚଏਈᩭ㉄ߩ᭴ᚑ߮᭴ᚑⷐ⚛ߩ↱᧪ߪ࿑ 1(p7) ߮ 1(p8㨪9)ߦ␜ߒߚߣ߅ࠅߢࠆޔߚ߹ޕ᭴ᚑⷐ⚛ߩႮၮ㈩ߪ MON 88017 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩᷝ⾗ᢱ 1 ߩ p32㨪33 ߮ p50㨪54 ߦ⸥タߐࠇߡࠆޕ ዏ⚵ᧄޔ឵߃࠻࠙ࡕࡠࠦࠪߦߪ㊁↢ဳ cp4 epsps ㆮવሶࠍᡷᄌߒߚ cp4 epsps ㆮવ ሶ߮㊁↢ဳ cry3Bb1 ㆮવሶࠍᡷᄌߒߚ cry3Bb1 ㆮવሶࠍዉߒߡ߅ࠅޔએਅߎ ߩㆮવሶࠍޟᡷᄌဳ cp4 epsps ㆮવሶޠ߮ޟᡷᄌဳ cry3Bb1 ㆮવሶࠆߔ⊒ޔޠ Ⱞ⊕⾰ࠍޟᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ޠ߮ޟᡷᄌဳ CP4 EPSPS Ⱞ⊕⾰ߣޠ⒓ߔࠆޕ MON810 ߩߦ↪ࠄࠇߚଏਈᩭ㉄ߩ᭴ᚑ߮᭴ᚑⷐ⚛ߩ↱᧪ߪ࿑ 2(p10) ߮ 2(p11)ߦ␜ߒߚߣ߅ࠅߢࠆޕ 5 cry1Ab ㆮવሶߩႮၮ㈩ߪ MON810 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩᷝ⾗ᢱ 1 ߩ p808(2 ᨎ⋡)ߩ FIGURE 1 ߦ␜ߒߚޕ 6 EcoR I 394 LB P-ract ori-V ract1 intron CTP2 ᡷᄌဳ cp4 epsps ROP PV-ZMIR39 EcoR I 3027 12368 bp NOS 3’ ori-322 P-e35S EcoR V 3842 Xba I 3963 aad wt CAB leader ᡷᄌဳ RB ract1 intron cry3Bb1 T-DNA 㗔ၞ Note: Sca I is a non-cutting enzyme tahsp17 3’ EcoR I 6510 ࿑ 1 MON 88017 ߩߦ↪ࠄࠇߚࡊࠬࡒ࠼ PV-ZMIR39 1 1 ᧄ࿑ߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߅ࠃ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 7 1 MON 88017 ߩߦ↪ࠄࠇߚࡊࠬࡒ࠼ PV-ZMIR39 ߩฦ᭴ᚑⷐ⚛↱᧪ ߮ᯏ⢻ 2 ᭴ᚑⷐ⚛ ↱᧪߮ᯏ⢻ ᡷᄌဳ cp4 epsps ㆮવሶࠞ࠶࠻ P-ract ract1 intron ࠗࡀ↱᧪ߩࠕࠢ࠴ࡦ 1 ㆮવሶߩࡊࡠࡕ࠲㗔ၞ⊛⋡ޕㆮવሶࠍ⊒ߐߖ ࠆ(ᢥ₂ 9)ޕ ࠗࡀࠕࠢ࠴ࡦㆮવሶߩࠗࡦ࠻ࡠࡦߩࠣࡦࠪࠗࡊࠬޕല₸ࠍ㜞ࠆߎߣ ߦࠃߞߡ⊛⋡ޔㆮવሶߩ⊒ࠍᵴᕈൻߐߖࠆ (ᢥ₂ 12)ޕ ࠪࡠࠗ࠽࠭࠽ߩ epsps ㆮવሶߩਛߢޔEPSPS Ⱞ⊕⾰ߩ N ᧃ┵ߦሽߔࠆ CTP2 ⪲✛ャㅍࡍࡊ࠴࠼ㇱಽࠍࠦ࠼ߔࠆႮၮ㈩(ᢥ₂ 13)⧐ޕ㚅ᣖࠕࡒࡁ㉄߇ วᚑߐࠇࠆ⪲✛߳ޔCP4 EPSPS Ⱞ⊕⾰ࠍャㅍߔࠆޕ Agrobacterium CP4 ⩶ᩣ↱᧪ߩ 5-ࠛࡁ࡞ࡇ࡞ࡆ࡞ࠪࠠࡒ㉄-3-ࡦ㉄วᚑ㉂ ᡷᄌဳ cp4 epsps ⚛ㆮવሶ(ᢥ₂ 14; ᢥ₂ 15)ޕᬀ‛ਛߢߩ⊒㊂ࠍ㜞ࠆߚޔCP4 EPSPS Ⱞ⊕⾰ߩᯏ⢻ᵴᕈࠍᄌᦝߔࠆߎߣߩߥࠃ߁ߦႮၮ㈩ߦᡷᄌࠍട߃ߚ߽ ߩߢ㉄ࡁࡒࠕޔ㈩ߦ㑐ߒߡߪ N ᧃ┵߆ࠄ 2 ⇟⋡ߩࡦ߇ࡠࠗࠪࡦߦᡷ ᄌߐࠇߚߩߺߢࠆޕᯏ⢻ߩ⚦ߦߟߡߪ p12㨪13 ߦ⸥タߒߚޕ Agrobacterium tumefaciens T-DNA ↱᧪ߩࡁࡄࡦวᚑ㉂⚛(NOS)ㆮવሶߩ NOS 3’ 3’㕖⠡⸶㗔ၞߢޔmRNA ߩォ౮ࠍ⚳⚿ߐߖ࡞࠾࠺ࠕࡐޔൻࠍ⺃ዉߔࠆ (ᢥ ₂ 16)ޕ ᡷᄌဳ cry3Bb1 ㆮવሶࠞ࠶࠻ P-e35S wt CAB leader ract1 intron ࠞࡈࡢࡕࠩࠗࠢ࠙ࠗ࡞ࠬ(CaMV)ߩࡊࡠࡕ࠲(ᢥ₂ 17)❱⚵ోޕਛ ߦ⋡⊛ㆮવሶࠍᕡᏱ⊛ߦ⊒ߐߖࠆᯏ⢻ࠍᜬߟޕ ࠦࡓࠡ⪲✛⚛ a/b ⚿วⰮ⊕⾰ߩ 5’ᧃ┵㕖⠡⸶࠳㗔ၞ⊛⋡ޕㆮવሶߩ⊒ ࠍᵴᕈൻߐߖࠆ(ᢥ₂ 18)ޕ ࠗࡀࠕࠢ࠴ࡦㆮવሶߩࠗࡦ࠻ࡠࡦߩࠣࡦࠪࠗࡊࠬޕല₸ࠍ㜞ࠆߎߣ ߦࠃߞߡ⊛⋡ޔㆮવሶߩ⊒ࠍᵴᕈൻߐߖࠆ (ᢥ₂ 12)ޕ Bacillus thuringiensis ߩᡷᄌߒߚ Cry3Bb1 Ⱞ⊕⾰ࠍࠦ࠼ߔࠆㆮવሶ(ᢥ₂ ᡷᄌဳ cry3Bb1 19)ޕMON 863 ߩᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߣߪࠕࡒࡁ㉄㈩߇৻ࡩᚲ⇣ߥࠅޔ 166 ⇟⋡ߩࠕࡒࡁ㉄㈩߇ MON 863 ߢߪࠣࠪࡦߢࠆߩߦኻߒߡ MON 88017 ߢߪࠕࠬࡄࠡࡦ㉄ߢࠆޕᯏ⢻⚦ߦߟߡߪ p13㨪14 ࠍෳᾖޕ tahsp 17 3’ 2 ࠦࡓࠡᾲ࡚ࠪ࠶ࠢⰮ⊕⾰ 17.3 ߩ 3’ᧃ┵㕖⠡⸶㗔ၞޕォ౮ࠍ⚳⚿ߐߖࡐޔ ࠕ࠺࠾࡞ൻࠍ⺃ዉߔࠆ(ᢥ₂ 20)ޕ ᧄߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 8 1 MON 88017 ߩߦ↪ࠄࠇߚࡊࠬࡒ࠼ PV-ZMIR39 ߩฦ᭴ᚑⷐ⚛↱᧪ ߮ᯏ⢻(⛯߈) 3 T-DNA 㗔ၞએᄖߩ᭴ᚑⷐ⚛ Ti ࡊࠬࡒ࠼ pTiT37 ߦ↱᧪ߔࠆ ဳࡦࡄࡁޔT-DNA ߩฝႺ⇇㈩ߩ DNA RB ᢿ ޕฝႺ⇇㈩ߪޔAgrobacterium tumefaciens ߆ࠄᬀ‛ࠥࡁࡓ߳ߩ T-DNA ߩવ㆐ߩ㓙ޔવ㆐ߩ㐿ᆎὐߣߒߡ↪ߐࠇࠆ(ᢥ₂ 21)ޕ Staphylococcus aureus ↱᧪ߩޔTn7 ࠕ࠺࠾࡞࠻ࡦࠬࡈࠚ(AAD)ࠍࠦ aad ࠼ߔࠆㆮવሶߢࠅࡦࠪࠗࡑࡁ࠴ࠢࡍࠬޔᚗߪࠬ࠻ࡊ࠻ࡑࠗࠪࡦ⠴ᕈࠍ ઃਈߔࠆ(ᢥ₂ 22)ޕ ori-322 ROP ori-V pBR322 ߆ࠄන㔌ߐࠇߚⶄ㐿ᆎ㗔ၞߢࠅޔE.coli ߦ߅ߡࡌࠢ࠲ߦ⥄ᓞ Ⴧᱺ⢻ࠍઃਈߔࠆ(ᢥ₂ 23)ޕ E. coli ਛߢߩࡊࠬࡒ࠼ߩࠦࡇᢙߩ⛽ᜬߩὑߦࡊࠗࡑ࠲ࡦࡄࠢ⾰ࠍᛥ ߔࠆࠦ࠺ࠖࡦࠣ㈩(ᢥ₂ 24)ޕ ᐢၞኋਥࡊࠬࡒ࠼ RK2 ߆ࠄන㔌ߐࠇߚⶄ㐿ᆎ㗔ၞߢࠅޔAgrobacterium tumefaciens ߦ߅ߡࡌࠢ࠲ߦ⥄ᓞჇᱺ⢻ࠍઃਈߔࠆ(ᢥ₂ 25)ޕ Ti ࡊࠬࡒ࠼ pTil5955 ߦ↱᧪ߔࠆᏀႺ⇇㈩ߩ DNA ᢿ ޕᏀႺ⇇㈩ߪޔ LB T-DNA ߇ Agrobacterium tumefaciens ߆ࠄᬀ‛ࠥࡁࡓ߳વ㆐ߐࠇࠆ㓙ߩ⚳⚿ὐ ߢࠆ(ᢥ₂ 26)ޕ 3 ᧄߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 9 㫃㪸㪺㩷 cry1Ab 㫃㪸㪺㩷 ᡷᄌဳ㩷 㪺㫇㪋㩷㪼㫇㫊㫇㫊㩷 ࿑ 2 MON810 ߩߦ↪ߚࡊࠬࡒ࠼ PV-ZMBK07 ߮ PV-ZMGT10 4 ᧄ࿑ߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 10 4 2 MON810 ߩߦ↪ࠄࠇߚࡊࠬࡒ࠼ PV-ZMBK07 ߮ PV-ZMGT10 ߩฦ᭴ ᚑⷐ⚛↱᧪߮ᯏ⢻ 5 ᭴ᚑⷐ⚛ ↱᧪߮ᯏ⢻ cry1Ab ㆮવሶࠞ࠶࠻ E35S ࠞࡈࡢࡕࠩࠗࠢ࠙ࠗ࡞ࠬ(CaMV)ߩ 35S ࡊࡠࡕ࠲(ᢥ₂ 17)߮ੑ㊀ࠛࡦ ࡂࡦࠨ㗔ၞࠍᜬߟ(ᢥ₂ 27)ޕ hsp70 intron ࠻࠙ࡕࡠࠦࠪߩᾲࠬ࠻ࠬⰮ⊕⾰ㆮવሶߩࠗࡦ࠻ࡠࡦޕhsp70 intron ߪᬀ‛ߦ߅ߌ ࠆᄖ᧪ㆮવሶߩ⊒㊂ࠍ㜞ࠆߚߦ↪ࠄࠇࠆ(ᢥ₂ 28)ޕ cry1Ab ფਛߦሽߔࠆ Bacillus thuringiensis subsp. krustaki HD-1 ᩣߩ Cry1Ab Ⱞ⊕⾰ࠍࠦ ࠼ߔࠆㆮવሶ(ᢥ₂ 29)ޕᯏ⢻ߩ⚦ߦߟߡߪ p14㨪15 ߦ⸥タߒߚޕ NOS 3’ Agrobacterium tumefaciens T-DNA ↱᧪ߩࡁࡄࡦวᚑ㉂⚛(NOS)ㆮવሶߩ 3’㕖⠡ ⸶㗔ၞߢޔmRNA ߩォ౮ࠍ⚳⚿ߐߖ࡞࠾࠺ࠕࡐޔൻࠍ⺃ዉߔࠆ (ᢥ₂ 16)ޕ ᡷᄌဳ cp4 epsps ㆮવሶࠞ࠶࠻(ᝌㆮવሶߩ⸃ᨆߩ⚿ᨐޔMON810 ਛߦߪᝌߐࠇߡߥ߆ߞߚ) E35S ࠞࡈࡢࡕࠩࠗࠢ࠙ࠗ࡞ࠬ(CaMV)ߩ 35S ࡊࡠࡕ࠲(ᢥ₂ 17)߮ੑ㊀ࠛࡦ ࡂࡦࠨ㗔ၞࠍᜬߟ(ᢥ₂ 27)ࠍᜬߟޕ hsp70 intron ࠻ ࠙ ࡕ ࡠ ࠦ ࠪ ߩ ᾲ ࠬ ࠻ ࠬ Ⱞ ⊕ ⾰ (heat shock protein) ㆮ વ ሶ ߩ ࠗ ࡦ ࠻ ࡠ ࡦ ޕ hsp70intron ߪᬀ‛ߦ߅ߌࠆᄖ᧪ㆮવሶߩ⊒㊂ࠍ㜞ࠆߚߦ↪ࠄࠇࠆ(ᢥ₂ 28)ޕ CTP2 ࠪࡠࠗ࠽࠭࠽ߩ epsps ㆮવሶߩਛߢޔEPSPS Ⱞ⊕⾰ߩ N ᧃ┵ߦሽߔࠆ⪲✛ ャㅍࡍࡊ࠴࠼ㇱಽࠍࠦ࠼ߔࠆ㈩ߢࠆ(ᢥ₂ 13)⧐ޕ㚅ᣖࠕࡒࡁ㉄߇วᚑߐࠇࠆ ⪲✛߳ࠍ⾰⊕Ⱞ⊛⋡ޔャㅍߔࠆޕ ᡷᄌဳ cp4 epsps Agrobacterium CP4 ⩶ᩣ↱᧪ߩ 5-ࠛࡁ࡞ࡇ࡞ࡆ࡞ࠪࠠࡒ㉄-3-ࡦ㉄วᚑ㉂ NOS 3’ ⚛ㆮવሶ(ᢥ₂ 14; ᢥ₂ 15)ޕ Agrobacterium tumefaciens T-DNA ↱᧪ߩࡁࡄࡦวᚑ㉂⚛(NOS)ㆮવሶߩ 3’㕖⠡ ⸶㗔ၞߢޔmRNA ߩォ౮ࠍ⚳⚿ߐߖ࡞࠾࠺ࠕࡐޔൻࠍ⺃ዉߔࠆ (ᢥ₂ 16)ޕ gox ㆮવሶࠞ࠶࠻(ᝌㆮવሶߩ⸃ᨆߩ⚿ᨐޔMON810 ਛߦߪᝌߐࠇߡߥ߆ߞߚ) E35S ࠞࡈࡢࡕࠩࠗࠢ࠙ࠗ࡞ࠬ(CaMV)ߩ 35S ࡊࡠࡕ࠲(ᢥ₂ 17)߮ੑ㊀ࠛࡦ ࡂࡦࠨ㗔ၞࠍᜬߟ(ᢥ₂ 27)ޕ hsp70 intron ࠻࠙ࡕࡠࠦࠪߩᾲࠬ࠻ࠬⰮ⊕⾰ㆮવሶߩࠗࡦ࠻ࡠࡦޕhsp70intron ߪᬀ‛ߦ߅ߌࠆ ᄖ᧪ㆮવሶߩ⊒㊂ࠍ㜞ࠆߚߦ↪ࠄࠇࠆ(ᢥ₂ 28) ࠪࡠࠗ࠽࠭࠽↱᧪ߩ rubisco ߩ small subunit 1A ㆮવሶߩਛߢޔrubisco small subunit 1A ߩ㧺ᧃ┵ߦሽߔࠆ⪲✛ャㅍࡍࡊ࠴࠼ㇱಽࠍࠦ࠼ߔࠆ㈩ߢࠆ(ޕᢥ₂ 30)⧐ޕ㚅ᣖࠕࡒࡁ㉄߇วᚑߐࠇࠆ⪲✛߳ࠍ⾰⊕Ⱞ⊛⋡ޔャㅍߔࠆޕ gox Achromobacter sp. strain LBAA ߩࠣࡎࠨ࠻ಽ⸃㉂⚛(glyphosate oxidoreductase; gox) ↱᧪ߩᄌ⇣ v247 ߩ C ᧃ┵ࠍࠦ࠼ߔࠆ㈩(ᢥ₂ 31)ޕGOX Ⱞ⊕⾰ߦࠃࠅ ࠣࡎࠨ࠻߇ಽ⸃ߐࠇࠆޕ NOS 3’ Agrobacterium tumefaciens T-DNA ↱᧪ߩࡁࡄࡦวᚑ㉂⚛(NOS)ㆮવሶߩ 3’㕖⠡⸶㗔 ၞߢޔmRNA ߩォ౮ࠍ㓸⚿ߐߖ࡞࠾࠺ࠕࡐޔൻࠍ⺃ዉߔࠆ (ᢥ₂ 16)ޕ ߘࠇએᄖߩ᭴ᚑⷐ⚛(PV-ZMBK07 ߮ PV-ZMGT10 ߦㅢ)(MON810 ਛߦߪᝌߐࠇߡߥ߆ߞߚ) lac Ǫ-D-ࠟࠢ࠻ࠪ࠳ߪ Lac Ⱞ⊕⾰ߩㇱಽ⊛ࠦ࠼㈩(ᢥ₂ 32)ޕǪ-ࠟࠢ࠻ ࠪ࠼ࠍಽ⸃ߒߡǪ-ࠟࠢ࠻ࠬࠍ↢ᚑߔࠆޕ ori-pUC ᄢ⣺⩶ࡊࠬࡒ࠼ pUC (ᢥ₂ 33)ߩⶄ㐿ᆎ㗔ၞࠍࠣࡔࡦ࠻ޕᄢ⣺⩶ࡊࠬ ࡒ࠼ߩⶄࠍ㐿ᆎߔࠆޕ nptII ේᩭ↢‛ߩ࠻ࡦࠬࡐ࠱ࡦ Tn5 ࠃࠅන㔌ߐࠇߚㆮવሶߢࡎࠬࡎࡦࠪࠗࡑࠝࡀޔ CTP 1 ࠻ࡦࠬࡈࠚ II ࠍࠦ࠼ߔࠆߩߎޕㆮવሶ߇ᓸ↢‛ౝߢ⊒ߐࠇࠆߣࠞ ࠽ࡑࠗࠪࡦ⠴ᕈ߇ઃਈߐࠇޔᒻ⾰ォ឵ߩㆬᛯࡑࠞߣߒߡߊ(ᢥ₂ 34)ޕ 5 ᧄߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 11 ࡠ ᭴ᚑⷐ⚛ߩᯏ⢻ MON 88017 ߩߦ↪ࠄࠇߚଏਈᩭ㉄ߩ᭴ᚑⷐ⚛ߩᯏ⢻ߪ 1(p8㨪9)ߦ␜ߒ ߚޕ MON 810 ߩߦ↪ࠄࠇߚଏਈᩭ㉄ߩ᭴ᚑⷐ⚛ߩᯏ⢻ߪ 2(p11)ߦ␜ߒߚޕ ޣᡷᄌဳ cp4 epsps ㆮવሶޤ ࠣࡎࠨ࠻ߪޔ㕖ㆬᛯ⊛ߥ㒰⨲ߢࠆ࠙ࡦ࠼ࠕ࠶ࡊߩലᚑಽߢ⧐ޔ㚅 ᣖࠕࡒࡁ㉄ߩ↢วᚑ⚻〝ߢࠆࠪࠠࡒ㉄วᚑ⚻〝ਛߩ㉂⚛ߩ৻ߟߢࠆ 5-ࠛࡁ ࡞ࡇ࡞ࡆ࡞ࠪࠠࡒ㉄-3-ࡦ㉄วᚑ㉂⚛(EPSPS)(E.C.2.5.1.19)ߣ․⇣⊛ߦ⚿วߒߡ ߘߩᵴᕈࠍ㒖ኂߔࠆ(ᢥ₂ 35; ᢥ₂ 36)ߚߩߘޕᬀ‛ߪࠣࡎࠨ࠻ࠍಣℂߔࠆ ߣ EPSPS ߇㒖ኂߐࠇࠆߎߣߦࠃࠅⰮ⊕⾰วᚑߦᔅ㗇ߩ⧐㚅ᣖࠕࡒࡁ㉄ࠍวᚑߢ߈ ߥߊߥࠅᨗࠇߡߒ߹߁ޕᡷᄌဳ cp4 epsps ㆮવሶߪ㒰⨲ࠣࡎࠨ࠻ߦ㜞⠴ᕈ ࠍᜬߟᡷᄌဳ CP4 EPSPS Ⱞ⊕⾰ࠍ⊒ߔࠆޕᡷᄌဳ cp4 epsps ㆮવሶߦࠃߞߡ↥ ↢ߐࠇࠆᡷᄌဳ CP4 EPSPS Ⱞ⊕⾰ߪ࠻ࠨࡎࠣޔሽਅߢ߽ᵴᕈ㒖ኂࠍฃߌߥ ߚ⚿ޔᨐߣߒߡᧄⰮ⊕⾰ࠍ⊒ߔࠆ⚵឵߃ᬀ‛ߢߪࠪࠠࡒ㉄วᚑ߇ᱜᏱߦᯏ ⢻ߒߡ↢⢒ߔࠆߎߣ߇ߢ߈ࠆޕ ዏޔEPSPS ߪᬀ‛߿ᓸ↢‛ߦ․ߩ⧐㚅ᣖࠕࡒࡁ㉄ࠍ↢วᚑߔࠆߚߩࠪࠠࡒ ㉄⚻〝ࠍ⸅ᇦߔࠆ㉂⚛ߩ৻ߟߢࠅޔᬀ‛ਛߢߪ⪲✛߹ߚߪ⦡⚛ߦሽߔࠆ (ᢥ₂ 37)ߪ〝⚻㉄ࡒࠠࠪޕᬀ‛ߩ࿕ቯߔࠆ⚛ߩ 5 ಽߩ 1 ߦ㑐ਈߔࠆߣ⠨߃ࠄࠇ ࠆ㊀ⷐߥઍ⻢⚻〝ߢࠆ(ᢥ₂ 38; ᢥ₂ 36)৻╙ߩߘޔߪ〝⚻ᧄޕᲑ㓏ߦ㑐ਈߔࠆ 3- ࠺ ࠝ ࠠ ࠪ -D- ࠕ ࡆ ࡁ - ࡋ ࡊ ࠷ ࡠ ࡦ ㉄ -7- ࡦ ㉄ (3-deoxy-D-arabino-heptulonate-7-phosphate, DAHP)วᚑ㉂⚛ߦࠃߞߡ⺞▵ࠍฃߌߡ ᓮߐࠇࠆ߇ޔDAHP ߆ࠄ EPSPS ߇⸅ᇦߔࠆ 5-ࠛࡁ࡞ࡇ࡞ࡆ࡞ࠪࠠࡒ㉄ 3 ࡦ ㉄(EPSP)ߩ↢ᚑࠍ⚻ߡࠦࠬࡒ㉄߇↢ᚑߐࠇࠆ߹ߢߩᲑ㓏ߢߪޔਛ㑆ઍ⻢‛⾰߿ ᦨ⚳↢ᚑ‛ߦࠃߞߡ㒖ኂߐࠇߚࠅᛥߐࠇࠆน⢻ᕈ߇ᭂߡૐߎߣ߇ࠄ߆ߦ ߐࠇߡࠆ(ᢥ₂ 39; ᢥ₂ 40) ߪߣߎߩߎޕEPSPS ߇ᧄ⚻〝ߦ߅ߌࠆᓞㅦ㉂⚛ߢߪ ߥߎߣࠍ␜ໂߒߡ߅ࠅޔᓥߞߡޔEPSPS ᵴᕈ߇Ⴧᄢߒߡ߽‛↥⚳ᦨߩ〝⚻ᧄޔ ߢࠆ⧐㚅ᣖࠕࡒࡁ㉄ߩỚᐲ߇㜞߹ࠆߎߣߪߥߣ⠨߃ࠄࠇߡࠆޕታ㓙ߦޔㅢ Ᏹߩ 40 ߩ EPSPS ࠍ↢ᚑߔࠆᬀ‛⚦⢩ߦ߅ߡ⧐ޔ㚅ᣖࠕࡒࡁ㉄߇ㆊߦวᚑ ߐࠇߥߎߣ߇ႎ๔ߐࠇߡ߅ࠅ(ᢥ₂ 41)ޔട߃ߡߦߢ߹ࠇߎ߇␠࠻ࡦࠨࡦࡕޔ ຠൻߒߚ㒰⨲࠙ࡦ࠼ࠕ࠶ࡊ⠴ᕈ‛(࠳ࠗ࠭)ࠪࠦࡠࡕ࠙࠻ޔ࠲ࡢޔࡀ࠲࠽ޔ ߩ㘩ຠ/㘺ᢱోᕈߩ⹏ଔߩㆊ⒟ߢ⚵ࠄࠇߘޔ឵߃‛⒳ሶਛߩࠕࡒࡁ㉄⚵ᚑࠍ⺞ ߴߡ⧐ޔ㚅ᣖࠕࡒࡁ㉄㊂ߦరߩ㕖⚵឵߃‛ߣߩ㑆ߢ⋧㆑ߩߥߎߣ߇⏕ߐ ࠇߡࠆ(ᢥ₂ 42; ᢥ₂ 43; ᢥ₂ 44; ᢥ₂ 45) ߪߣߎߩࠄࠇߎޕEPSPS ߇ᧄ⚻〝ߦ ߅ߌࠆᓞㅦ㉂⚛ߢߪߥߎߣࠍᡰᜬߒߡࠆޔߚ߹ޕEPSPS ߪࡎࠬࡎࠛࡁ࡞ࡇ ࡞ࡆࡦ㉄ (PEP)ߣࠪࠠࡒ㉄-3-ࡦ㉄ (S3P)߆ࠄޔEPSP ߣήᯏࡦ㉄ (Pi)ࠍ↢ߕࠆ 12 นㅒᔕࠍ⸅ᇦߔࠆ㉂⚛ߢࠅ(ᢥ₂ 46)ߩࠄࠇߎޔၮ⾰ߣ․⇣⊛ߦᔕߔࠆߎߣ ߇⍮ࠄࠇߡࠆ(ᢥ₂ 47)ࠄࠇߎޕએᄖߦ໑৻ EPSPS ߣᔕߔࠆߎߣ߇⍮ࠄࠇߡ ࠆߩߪ S3P ߩ㘃ૃߢࠆࠪࠠࡒ㉄ߢࠆ߇ޔ ߘߩᔕᕈߪ S3P ߣߩᔕᕈߩ 200 ਁಽߩ 1 ߦߔ߉ߕ↢ޔౝߢၮ⾰ߣߒߡᔕߔࠆߣߪ⠨߃ࠄࠇߥޕ ԙᡷᄌဳ CP4 EPSPS Ⱞ⊕⾰߇ޔᣢ⍮ߩធ⸅ࠕ࡞ࠥࡦߣᯏ⢻㊀ⷐߥࠕࡒࡁ㉄ ㈩ࠍߔࠆ߆ߤ߁߆(ࠬࡌ࠲࠺ޔGenBank, EMBL, PIR, NRL3D, SwissProt) ࠍ↪ߡᲧセߒߚߣߎࠈޔᣢ⍮ࠕ࡞ࠥࡦߣ᭴ㅧ⊛ߦ㘃ૃᕈߩࠆ㈩ࠍߒ ߡߥ߆ߞߚޕ ޣᡷᄌဳ cry3Bb1 ㆮવሶޤ Ԙࠦ࠙࠴ࡘ࠙⋡ኂ⯻ᛶ᛫ᕈࠍઃਈߔࠆߚߩ⋡⊛ㆮવሶߢࠆᡷᄌဳ cry3Bb1 ㆮવሶߪޔფਛߦ᥉ㆉ⊛ߦሽߔࠆࠣࡓ㓁ᕈ⩶ߢࠆ Bacillus thuringiensis subsp. kumamotoensis ߦ↱᧪ߒࠆࠇߐ࠼ࠦޔᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߪ☨࿖ߩ࠻࠙ ࡕࡠࠦࠪᩱၭߩਥⷐࠦ࠙࠴ࡘ࠙⋡ኂ⯻ߢࠅߩࠪࠦࡠࡕ࠙࠻ޔᩮࠍ㘩ኂߔࠆ corn rootworm (Diabrotica sp.)( એਅ CRW ߣߔࠆ) ߦኻߔࠆᲕ⯻ᵴᕈࠍ␜ߔޕᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ࠍߚ B.t.⩶ߩ↥↢ߔࠆ B.t.Ⱞ⊕⾰ߪޔᮡ⊛⯻ߩਛ⣺⊹ߩ․ ⇣⊛ฃኈߣ⚿วߒߡ㓁ࠗࠝࡦㆬᛯ⊛ዊሹࠍᒻᚑߒ⚿ߩߘޔᨐޔᶖൻࡊࡠࠬࠍ 㒖ኂߒߡᲕ⯻ᵴᕈࠍ␜ߔ(ᢥ₂ 48; ᢥ₂ 49; ᢥ₂ 50)ޕ Cry3Bb1 Ⱞ⊕⾰ߩᲕ⯻ࠬࡍࠢ࠻ࡓߪᭂߡ⁜ߊ⋡࠙ࡘ࠴࠙ࠦޔ⯻⒳ߩਛߢ ࡂ ࡓ ࠪ ⑼ ߩ 2 ዻ (Leptinotarsa, Diabrotica) ߦ ಽ 㘃 ߐ ࠇ ࠆ Colorado potato beetle(Leptinotarsa decimlineata) (એਅ CPB ߣߔࠆ)ߣ CRW ߩߺߦኻߒߡᲕ⯻ᵴᕈࠍ ␜ߔ(MON 88017 ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩᷝ⾗ᢱ 3 ߩ p108 ߩ Table 1) ߩߎޕ2 ዻߩ⯻⒳ߣߩหዻㄭ✼⒳߇ࠊ߇࿖ߦ↢ᕷߒߡࠆߎߣߪߎࠇ߹ߢႎ๔ߐࠇߡ ߥߎߣ߇ᢥ₂⺞ᩏߦࠃࠅ␜ߐࠇߚ(MON 88017 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩᷝ ⾗ᢱ 3 ߩ p54 ߩ 10, ᢥ₂ 51)ޕ ߥ߅ޔᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߪ㊁↢ဳ Cry3Bb1 Ⱞ⊕⾰ߣᲧセߒߡޔ98%એߩ ⋧หᕈࠍߒߡࠆ(MON 88017 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠᷝ⾗ᢱ 2 ߩ p20 ߩ࿑ 5-2)⋡࠙ࡘ࠴࠙ࠦޕኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ MON 863 ߩᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߣ MON 88017 ߩᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߣߪࠕࡒࡁ㉄㈩߇৻ࡩᚲ⇣ߥࠅޔ166 ⇟⋡ ߩࠕࡒࡁ㉄㈩߇ MON 863 ߢߪࠣࠪࡦߢࠆߩߦኻߒߡᧄ⚵឵߃࠻࠙ࡕࡠࠦ ࠪߢߪࠕࠬࡄࠡࡦ㉄ߢࠆޕታ㓙ߦ߶႐ߦ߅ߌࠆᲕ⯻ലᨐߪ MON 88017 ߩᡷᄌ ဳ Cry3Bb1 Ⱞ⊕⾰ࠍ↪ߡⴕߞߡࠆޕ ԙᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰߇ᣢ⍮ߩធ⸅ࠕ࡞ࠥࡦߣᯏ⢻㊀ⷐߥࠕࡒࡁ㉄㈩ ࠍߔࠆ߆ߤ߁߆ࠍ࠺࠲ࡌࠬ(GenBank, EMBL, PIR, NRL3D, SwissProt)ࠍ↪ 13 ߡᲧセߒߚߣߎࠈޔᣢ⍮ࠕ࡞ࠥࡦߣ᭴ㅧ⊛ߦ㘃ૃᕈߩࠆ㈩ࠍߒߡ ߥ߆ߞߚޕ ޣcry1Ab ㆮવሶޤ ԘMON810 ߢ࠴࡚࠙⋡ኂ⯻ᛶ᛫ᕈࠍઃਈߔࠆߚߩ⋡⊛ㆮવሶߢࠆ cry1Ab ㆮ વሶߪޔ ფਛߦ᥉ㆉ⊛ߦሽߔࠆࠣࡓ㓁ᕈ⩶ߢࠆ Bacillus thuringiensis subsp. kurstaki ߦ↱᧪ߒ ࠆࠇߐ࠼ࠦޔCry1Ab Ⱞ⊕⾰ߪ☨࿖ߩ࠻࠙ࡕࡠࠦࠪᩱၭߩਥⷐ ࠴࡚࠙⋡ኂ⯻ߢࠆ European corn borer (Ostrinia nubilalis)ߦኻߔࠆᲕ⯻ᵴᕈࠍ ߔࠆ(Frankenhuzen, 1993)ޕEuropean corn borer ߩ㘩ኂㇱߪᬀ‛ㇱో⥸ߢ ࠆޕCry1Ab Ⱞ⊕⾰ࠍߚ B.t.⩶ߩ↥↢ߔࠆ B.t.Ⱞ⊕⾰ߪޔᮡ⊛⯻ߩਛ⣺⊹ ߩ․⇣⊛ฃኈߣ⚿วߒߡ㓁ࠗࠝࡦㆬᛯ⊛ዊሹࠍᒻᚑߒ⚿ߩߘޔᨐޔᶖൻࡊࡠ ࠬࠍ㒖ኂߒߡᲕ⯻ᵴᕈࠍ␜ߔ(ᢥ₂ 48; ᢥ₂ 50)ޕB.t.Ⱞ⊕⾰ߪ㉂⚛ᵴᕈࠍᜬߚߕޔ ኋਥߩઍ⻢♽ߣߪ⁛┙ߒߡᯏ⢻ߒߡࠆޕ Cry1Ab Ⱞ⊕⾰ߪ⋡࡚࠙࠴ޔ⯻ߦߩߺᲕ⯻ᵴᕈࠍ␜ߒ⋡࡚࠙࠴ޔએᄖߩ⯻ߦ ኻߒߡߪᲕ⯻ᵴᕈࠍᜬߚߥ ߩߎޔߚ߹ޕCry1Ab Ⱞ⊕⾰ߪ☨ޔ࿖ߩ࠻࠙ࡕࡠࠦ ࠪᩱၭߦ߅ߌࠆ㊀ⷐኂ⯻ߢࠆ࠴࡚࠙⋡ߩ European corn borer (Ostrinia nubilalis), Southwestern corn borer (Diatraea grandiosella), Southern cornstalk borer (Diatraea crambidoides), Sugarcane cornstalk borer (Diatraea saccharalis), Corn earworm (Helicoverpa zea), Fall armyworm (Spodoptera frugiperda), Stalk borer (Papaipema nebris)ߦኻߒߡᲕ⯻ᵴᕈࠍ␜ߔߎߣ߇⍮ࠄࠇߡࠆ(ᢥ₂ 52; ᢥ₂ 53; ᢥ₂ 54; ᢥ ₂ 55; ᢥ₂ 56)ߩࠄࠇߎޕౝޔO. nubilalis ߣหዻߩ O. furnacalis ߪᣣᧄߩ࠻࠙ࡕࡠ ࠦࠪᩱၭߦ߅ߌࠆਥⷐ࠴࡚࠙⋡ኂ⯻ߣߒߡ⍮ࠄࠇߡࠆ(ᢥ₂ 57)ޕ ԙCry1Ab Ⱞ⊕⾰߇ޔᣢ⍮ߩࠕ࡞ࠥࡦߣᯏ⢻㊀ⷐߥࠕࡒࡁ㉄㈩ࠍߔࠆ ߆ߤ߁߆(ࠬࡌ࠲࠺ޔGenBank, EMBL, PIR, NRL3D, SwissProt)ࠍ↪ߡᲧセߒ ߚߣߎࠈޔᣢ⍮ࠕ࡞ࠥࡦߣ᭴ㅧ⊛ߦ㘃ૃᕈߩࠆ㈩ࠍߒߡߥ߆ߞߚޕ ޣᡷᄌဳ cry3Bb1 ㆮવሶ+cry1Ab ㆮવሶޤ ᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰߇ዻߔࠆ Cry3 ࡈࠔࡒߣ Cry1Ab Ⱞ⊕⾰߇ዻߔࠆ Cry1A ࡈࠔࡒߪ⋡࠙ࡘ࠴࠙ࠦࠇߙࠇߘޔ⯻߮࠴࡚࠙⋡⯻ߣ߁⇣ߥࠆ ⋡ߦಽ㘃ߐࠇࠆ⯻⒳ߩᐜ⯻ߦኻߒߡ․⇣⊛ߦᲕ⯻ᵴᕈࠍ␜ߔߎߣ߇ޔ1960 ᐕ߆ ࠄ↢‛ㄘ⮎ߣߒߡ↪ߐࠇߡࠆ B.t.ߩ⍮߆ࠄ߽⍮ࠄࠇߡࠆޕ ߹ߚޔਔⰮ⊕⾰ߣ߽ᮡ⊛⯻ߩਛ⣺⊹⚦⢩ߩ․⇣⊛ฃኈߣ⚿วߒߡᶖൻࡊ ࡠࠬࠍ㒖ኂߔࠆߣ߁↪ᯏߪㅢߒߡࠆ߇⋡࡚࠙࠴ޔ⯻ߣࠦ࠙࠴ࡘ࠙ ⋡⯻ߩᐜ⯻ߦ߅ߌࠆਛ⣺ౝߩ pH ߪࠞ࡞ࠕࠇߙࠇߘޔᕈ(pH 10.5㨪pH 11.0)ߣ 14 ਛᕈ(pH 6.5㨪pH 7.0)ߢࠅޔਔⰮ⊕⾰ߪ⇣ߥࠆൻቇ᧦ઙߢߘࠇߙࠇᲕ⯻ᵴᕈࠍ⊒ ើߒߡࠆ(ᢥ₂ 58)ޕ ᦝߦޔMacIntosh ࠄߩႎ๔ߦࠃࠆߣ ޔ3(p16)ߦ␜ߒߚࠃ߁ߦ Cry1Ab Ⱞ⊕⾰ߣ หߓ Cry1A ࡈࠔࡒߦዻߔࠆ Cry1Ac Ⱞ⊕⾰ߣޔᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߣหߓ Cry3 ࡈࠔࡒߦዻߔࠆ Cry3Aa Ⱞ⊕⾰ߦኻߒߡᗵฃᕈࠍ␜ߐߥ㕖ᮡ⊛⯻ߪޔ ߎࠇࠄ 2 ߟߩ⇣ߥࠆࡈࠔࡒߦዻߔࠆ B.t.Ⱞ⊕⾰ࠍᷙวߒߡਈ߃ߚ႐วߢ߽ޔᄌ ࠊࠄߕ㕖ᗵฃᕈߩ߹߹ߢࠅޔ Cry1Ac Ⱞ⊕⾰ߣ Cry3A Ⱞ⊕⾰ߦหᤨߦ㔺ߐࠇࠆ ߎߣߦࠃࠆ⋧ਸ਼⊛ߥᓇ㗀ࠍฃߌߥߎߣ߇ႎ๔ߐࠇߡࠆ(ᢥ₂ 59)ޕᓥߞߡޔᓥ ᧪ߩ㔀⢒ᚑᴺࠍ↪ߡ㈩ߐࠇߚ㈩ᓟઍຠ⒳ߢࠆᧄࠬ࠲࠶ࠢ♽⛔࠻࠙ࡕࡠ ࠦࠪਛߦߪⷫ♽⛔↱᧪ߩ Cry1Ab Ⱞ⊕⾰ߣᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰߇⊒ߒߡࠆ ߇߇ࠄࠇߘޔߩ㕖ᮡ⊛⯻ߦኻߒߡ⋧ਸ਼ലᨐ⊛ߦᲕ⯻ᵴᕈࠍ␜ߔน⢻ᕈߪ߈ ࠊߡૐߣ⠨߃ࠄࠇߚޕ 15 3 Cry1 ߮ Cry3 Ⱞ⊕⾰ߩ⇣ߥࠆ⯻⒳ߦኻߔࠆᲕ⯻ᵴᕈࠍ⺞ᩏߒߚ⚿ᨐ (MacIntosh ࠄߩ⺰ᢥ╬ࠍ߽ߣߦᚑߒߚ)ޕ Cockroach (Blatella germanica) Alfalfa weevil (Hypera postica) Cotton boll weevil (Antonomus grandes) Horseradish flea beetle (Phyllotreta armoraciae) Southern corn rootworm (Diabrotica undecimpunctata howardii) Japanese beetle (Popilla japonica) Colorado potato beetle (Leptinotarsa decemlineata) Mosquito (Aedes aegypti) Green peach aphid (Myzus persicae) Termite (Reticulitermes flavipes) Beet armyworm (Spodoptera exigua) Black cutworm (Agrotis ipsilon) Cabbage looper (Trichoplusia ni) Corn earworm (Heliothis zea) European corn borer (Ostrinia nubilialis) Tobacco budworm (Heliothis virescens) Tobacco hornworm (Manduca sexta) Spider mite (Tetranychus urticae) Cry1Ab (1 Cry1Ac (2 Cry3A (3 Cry1Ac + Cry3A (4 - - - - - - - - - - - - - - - - - - - - - - - - - - + + - - - - - - - - - - - - + + - + + + - + + + - + + + - + + + - + + + - + + + - + - - - - Cry1Ab Ⱞ⊕⾰ߩỚᐲ߇ੱᎿ㘺ᢱਛߢ 50Ǵg/ml ߦߥࠆࠃ߁ߦ⺞ᢛߒߡ⯻⒳ߦਈ߃ߚޕ Cry1Ac Ⱞ⊕⾰ߩỚᐲ߇ੱᎿ㘺ᢱਛߢ 50Ǵg/ml ߦߥࠆࠃ߁ߦ⺞ᢛߒߡ⯻⒳ߦਈ߃ߚޕ Cry3A Ⱞ⊕⾰ߩỚᐲ߇ੱᎿ㘺ᢱਛߢ 500Ǵg/ml ߦߥࠆࠃ߁ߦ⺞ᢛߒߡ⯻⒳ߦਈ߃ߚޕ Cry1Ac Ⱞ⊕⾰ߩỚᐲ߇ੱᎿ㘺ᢱਛߢ 50Ǵg/mlޔ Cry3A Ⱞ⊕⾰߇ 500Ǵg/ml ߦߥࠆࠃ߁ߦ⺞ᢛߒߡ⯻⒳ߦਈ߃ߚޕ 㧗 ᬌቯᐜ⯻ߩᱫ₸߇ 25%એߢߞߚ⹜㛎 㧙 ᬌቯᐜ⯻ߩᱫ₸߇ 25㧑એਅߢߞߚ⹜㛎 1- 2- 3- 4- 16 (2) ࡌࠢ࠲ߦ㑐ߔࠆᖱႎ ࠗ ฬ⒓߮↱᧪ MON 88017 ߩߦ↪ࠄࠇߚࡌࠢ࠲ߪޔᄢ⣺⩶(Escherichia coli)↱᧪ߩࡊ ࠬࡒ࠼ pBR322 ߥߤࠍ߽ߣߦ᭴▽ߐࠇߚ(ᢥ₂ 23)ޕMON 810 ߩߦ↪ࠄࠇߚ ࡌࠢ࠲ߪޔᄢ⣺⩶(Escherichia coli)↱᧪ߩࡊࠬࡒ࠼ pUC119 ߥߤࠍ߽ߣߦ᭴▽ ߐࠇߚ(ᢥ₂ 33)ޕ ࡠ ․ᕈ MON 88017 ߩߦ↪ࠄࠇߚ PV-ZMIR39 ߩోႮၮᢙߪ 12,368bp ߢࠅޔᄢ ⣺⩶ߦ߅ߌࠆ᭴▽ࡌࠢ࠲ߩㆬᛮࡑࠞㆮવሶߣߒߡࠬࡍࠢ࠴ࡁࡑࠗࠪࡦࠆ ߪࠬ࠻ࡊ࠻ࡑࠗࠪࡦ⠴ᕈࠍઃਈߔࠆߚߩ aad ㆮવሶࠍᜬߟ(ᢥ₂ 22)ࡌᧄޕ ࠢ࠲ߩᗵᨴᕈߪ⍮ࠄࠇߡߥޕ MON810 ߩߦ↪ࠄࠇߚ PV-ZMBK07 ߮ PV-ZMGT10 ߩోႮၮᢙߪߘࠇ ߙࠇ 7,794 bpޔ9,427 bp ߢࠅޔᄢ⣺⩶ߦ߅ߌࠆ᭴▽ࡌࠢ࠲ߩㆬᛮࡑࠞㆮવ ሶߣߒߡ࠻ࡦࠬࡐ࠱ࡦ Tn5 ↱᧪ߩࠞ࠽ࡑࠗࠪࡦ㧛ࡀࠝࡑࠗࠪࡦ⠴ᕈㆮવሶ(nptII ㆮવሶ)ࠍᜬߟߩ࠲ࠢࡌᧄޕᗵᨴᕈߪ⍮ࠄࠇߡߥޕ (3) ㆮવሶ⚵឵߃↢‛╬ߩ⺞ᣇᴺ ࠗ ኋਥౝߦ⒖ߐࠇߚᩭ㉄ోߩ᭴ᚑ MON 88017 ߩߦߪޔ⸥ߩ aad ㆮવሶࠍᜬߟ pBR322 ߥߤࠍరߦࠬࡊޔ ࡒ࠼ࡌࠢ࠲PV-ZMIR39 ࠍ᭴▽ߒߚޕ ኋਥౝߦ⒖ߐࠇߚᧄࡌࠢ࠲ਛߩ T-DNA 㗔 ၞ ߪ ޔᡷ ᄌ ဳ cp4 epsps ㆮ વ ሶ ࠞ ࠶ ࠻ [P-ract]-[ract1 intron]-[CTP2]-[cp4 epsps]-[NOS 3’]߮ᡷᄌဳ cry3Bb1 ㆮવሶࠞ࠶࠻[P-e35S]-[wt CAB leader]-[ract1 intron]-[cry3Bb1]-[tahsp17 3’]ߢ᭴ᚑߐࠇࠆޕዏޔPV-ZMIR39 ߩࡊࠬࡒ࠼ࡑ࠶ࡊߪޔ p6 ߩ࿑ 1 ߦ⸥タߒߚޕ MON810 ߩߦߪޔ ⸥ߩ nptII ㆮવሶࠍᜬߟ pUC119 ↱᧪ߩࡌࠢ࠲ࠍరߦޔ Ԙcry1Ab ㆮવሶࠞ࠶࠻([E35S]-[hsp70 intron]-[cry1Ab]-[NOS3’])ࠍㅪ⚿ߒߚࡊ ࠬࡒ࠼ PV-ZMBK07ߦ߮ࠄߥޔԙᡷᄌဳ cp4 epsps ㆮવሶࠞ࠶࠻([E35S]-[hsp70 intron]-[CTP2]-[cp4 epsps]-[NOS 3’]) ߣ gox ㆮ વ ሶ ࠞ ࠶ ࠻ ([E35S]-[hsp70 intron]-[CTP1]-[gox]-[NOS 3’])ࠍㅪ⚿ߒߚࡊࠬࡒ࠼ PV-ZMGT10 ࠍ᭴▽ߒ ߩߎޔ2 ߟࠍࡌࠢ࠲ߣߒߡ↪ߚޕዏޔPV-ZMBK07 ߮ PV-ZMGT10 ߩࡊࠬࡒ࠼ࡑ 17 ࠶ࡊߪޔp9 ߩ࿑ 2 ߦ⸥タߒߚޕ ࡠ ኋਥౝߦ⒖ߐࠇߚᩭ㉄ߩ⒖ᣇᴺ MON 88017 ߩߢߪ࠲ࠢࡌ࠼ࡒࠬࡊޔPV-ZMIR39 ਛߩ T-DNA 㗔ၞࠍࠕ ࠣࡠࡃࠢ࠹࠙ࡓᴺߦࠃࠅ࠻ࡦ࠺ޔ⒳ߦಽ㘃ߐࠇࠆຠ⒳ߢࠆ A x HiII ߦዉߒ ߚޕ MON 810 ߩߢߪ ࠼ࡒࠬࡊޔPV-ZMBK07 ߮ PV-ZMGT10 ߩᷙว‛ࠍࡄ ࠹ࠖࠢ࡞ࠟࡦᴺߦࠃߞߡ࠻ࡦ࠺ޔ⒳ߦಽ㘃ߐࠇࠆ࠻࠙ࡕࡠࠦࠪ⥄ᱺ♽⛔ A188 X B73 ߩ F2 ઍߦዉߒߚޕ ࡂ ㆮવሶ⚵឵߃↢‛╬ߩ⢒ᚑߩ⚻ㆊ ޣMON 88017 ߩ⢒ᚑߩ⚻ㆊޤ ԘMON 88017 ߩ㐿⊒ߪ 1999 ᐕ߆ࠄᆎ߹ߞߚࡓ࠙࠹ࠢࡃࡠࠣࠕޕᴺߦࠃࠅࡊ ࠬࡒ࠼ࡌࠢ࠲PV-ZMIR39 ਛߩ T-DNA 㗔ၞࠍ A x HiII ߦዉߒߚᓟࡎࠣޔ ࠨ࠻ࠍၭߢᒻ⾰ォ឵ࠞ࡞ࠬࠍㆬᛮߒߡౣ↢ࠍᓧߡޔᡷᄌဳ Cry3Bb1 Ⱞ⊕⾰ߩ⊒ࠍ ELISA ಽᨆߦࠃߞߡᬌቯߒ࠻ࠨࡎࠣޔ⠴ᕈߣኂ⯻ᛶ ᛫ᕈߩઃਈߐࠇߚ♽⛔ࠍㆬᛮߒߚޕ ԙߎߩ㓙ޔᒻ⾰ォ឵ࠞ࡞ࠬࠍࠞ࡞ࡌ࠾ࠪࡦࠍၭߢၭ㙃ߒߚᓟࠄࠇߎޔ ᛫↢‛⾰ࠍ߹ߥౣ↢ၭߦ⒖ߒߡၭ㙃ߔࠆߎߣߦࠃߞߡ࠙࠹ࠢࡃࡠࠣࠕޔ ࡓߩᱷሽᕈ߇ߥߎߣࠍ⏕ߒߡࠆ(ᢥ₂ 60)ޕ Ԛ2000㨪2001 ᐕߦ߆ߌߡᑧߴ 169 ࡩᚲߩ߶႐⹜㛎ࠍⴕ⚳ᦨޔຠൻ♽⛔ࠍㆬ ᛮߔࠆߣߣ߽ߦߩߘޔⅣႺోᕈࠍ⹏ଔߒߚ(⹜㛎ߦ↪ߚ♽⛔ߦߟߡߪ p20 ߩ ࿑ 3 ࠍෳᾖ)ޕ MON 88017 ߩࠊ߇࿖ߦ߅ߌࠆน⁁ᴫߪએਅߩㅢࠅߢࠆޕ 2003 ᐕ 4 2004 ᐕ 8 ㄘᨋ᳓↥⋭ࠃࠅޟㄘᨋ᳓↥ಽ㊁╬ߦ߅ߌࠆ⚵឵߃ߩ↪ߩߚ ߩᜰ㊎ߦޠၮߠ߈ޔᣣᧄߢߩᩱၭߦߟߡޔᜰ㊎߳ߩㆡวᕈ ߇⏕ߐࠇߚޕ ㄘᨋ᳓↥⋭߮ⅣႺ⋭ࠃࠅㆮવሶ⚵ߺ឵߃↢‛╬ߩ↪╬ߩⷙ ߦࠃࠆ↢‛ߩᄙ᭽ᕈߩ⏕ߦ㑐ߔࠆᴺᓞߦၮߠߊ╙৻⒳↪ ⷙ⒟(㘩↪߹ߚߪ㘺ᢱ↪ߦଏߔࠆߚߩ↪ޔᩱၭޔടᎿޔ▤ޔ ㆇ៝߮ᑄ᫈ਗ߮ߦߎࠇࠄߦઃ㓐ߔࠆⴕὑߦߟߡ)ߩክᩏࠍ ⚳ੌߒߚޕ 18 ޣMON810 ߩ⢒ᚑߩ⚻ㆊޤ ԘPV-ZMBK07 ߮ PV-ZMGT10 ࠍዉߒߚࠞ࡞ࠬࠍޔ2,4-D ࠍ⚵❱ၭ㙃ၭ ߢߒ߫ࠄߊ↢⢒ߐߖߚᓟࠍ࠻ࠨࡎࠣޔၭߢ⚵឵߃ࠍㆬᛮߒޔㆬ ᛮߐࠇߚࠞ࡞ࠬ߆ࠄౣ↢ࠍᓧߡ Cry1Ab Ⱞ⊕⾰ߩ⊒ࠍ⸃ᨆߒߚޕዏޔ MON810 ߩࠨࠩࡦࡉࡠ࠶࠻ߦࠃࠆᝌㆮવሶ⸃ᨆߩ⚿ᨐޔnptII ㆮવሶޔᡷᄌဳ cp4 epsps ㆮવሶޔgox ㆮવሶߩ⊒ࠞ࠶࠻ߪሽߒߥߎߣ߇⏕ߐࠇߚ(MON810 ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩᷝ⾗ᢱ 2 ߩ p22 ߩ࿑ 4)ޕMON810 ߦᡷᄌဳ cp4 epsps ㆮવሶ߇ᝌߐࠇߡߥ߆ߞߚߦ߽߆߆ࠊࠄߕࠣࡎࠨ࠻ߢㆬᛮߐࠇߚߩߪޔ ౣಽൻ(R0)ߩᰴઍ(BC0F1)ߢᝌㆮવሶߦ㑐ߒߡಽ㔌߇߈ߚߚߢࠆ น⢻ᕈ߇⠨߃ࠄࠇߚޔߒ߆ߒޕMON810 ߪኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪߣߒߡㆬᛮ߇ ㅴࠄࠇޔౣಽൻߩᰴઍߢࠣࡎࠨ࠻ᬌቯ߮ࠨࠩࡦࡉࡠ࠶࠻ಽᨆߪⴕ ࠊࠇߥ߆ߞߚߚޔℂ↱ߪ․ቯߐࠇߥ߆ߞߚޕ ԙMON810 ߪࡄ࠹ࠖࠢ࡞ࠟࡦᴺߦࠃߞߡ DNA ᢿ ࠍዉߒߡࠆߚࠕޔ ࠣࡠࡃࠢ࠹࠙ࡓߩᱷሽᕈߩ⏕ߪⴕࠊߥ߆ߞߚޕ Ԛ1992 ᐕࠃࠅ♽⛔ㆬᛮߩ⹏ଔࠍ㐿ᆎߒޔ1993㨪1995 ᐕߦ߆ߌߡ႐⹜㛎ࠍⴕߞ ߡߦ⊛⚳ᦨޔఝ⦟♽⛔ࠍㆬᛮߒߚޔߡߒߘޕ1994 ᐕߦⴕߞߚ☨࿖ 6 ࡩᚲߩ႐⹜ 㛎ߦ߅ߡߩ⛔♽ᧄޔᒻᘒ߮↢⢒․ᕈߥߤߦߟߡ⺞ᩏࠍⴕ߁ߣߣ߽ߦޔ Cry1Ab Ⱞ⊕⾰ߩ⊒߮ᝌㆮવሶߩಽᨆ╬ࠍⴕߞߚ(⹜㛎ߦ↪ߚ♽⛔ߦߟ ߡߪ p20 ߩ࿑ 4 ࠍෳᾖ)ޕ ߘࠇࠄߩ⚿ᨐߦၮߠߡޔ ☨࿖ߢᔅⷐߥนࠍฃߌߡ 1997 ᐕ߆ࠄ৻⥸ᬺᩱၭ߇ᆎࠄࠇߡࠆޕ MON810 ߩࠊ߇࿖ߦ߅ߌࠆน⁁ᴫߪએਅߩㅢࠅߢࠆޕ 1996 ᐕ 10 ㄘᨋ᳓↥⋭ࠃࠅޟㄘᨋ᳓↥ಽ㊁╬ߦ߅ߌࠆ⚵឵߃ߩ↪ߩߚ ߩᜰ㊎ߦޠၮߠ߈ޔᣣᧄ߳ߩャ(ടᎿ↪߮㘺ᢱ↪ߣߒߡ ߩ↪)ߦߟߡޔᜰ㊎߳ߩㆡวᕈ߇⏕ߐࠇߚޕ 1997 ᐕ 5 ෘ↢ഭ⋭(ᒰᤨෘ↢⋭)ࠃࠅ⚵ޟ឵߃ DNA ᛛⴚᔕ↪㘩ຠ㘩 ຠᷝട‛ߩోᕈ⹏ଔᜰ㊎ߦޠၮߠ߈ޔ㘩ຠ↪ߣߒߡߩో ᕈนࠍฃߌߚޕ 1997 ᐕ 6 ㄘᨋ᳓↥⋭ࠃࠅ⚵ޟ឵߃↪㘺ᢱߩోᕈ⹏ଔᜰ㊎ 6 ߩ(2)ޠ ߦၮߠ߈ޔ㘺ᢱ↪ߣߒߡߩోᕈนࠍฃߌߚޕ 2001 ᐕ 3 ෘ↢ഭ⋭ࠃࠅ⚵ޟ឵߃ DNA ᛛⴚᔕ↪㘩ຠ߮ᷝട‛ߩో ᕈክᩏߩᚻ⛯ߦޠၮߠ߈ޔ㘩ຠ↪ߣߒߡߩోᕈ⏕ࠍฃߌ ߚޕ 2003 ᐕ 3 ㄘᨋ᳓↥⋭ࠃࠅ⚵ޟ឵߃ DNA ᛛⴚᔕ↪㘺ᢱ߮㘺ᢱᷝട‛ߩ ోᕈߦ㑐ߔࠆ⏕ߩᚻ⛯ߦޠၮߠ߈ޔ㘺ᢱ↪ߣߒߡߩో ᕈ⏕ࠍฃߌߚޕ 2003 ᐕ 4 ㄘᨋ᳓↥⋭ࠃࠅޟㄘᨋ᳓↥ಽ㊁╬ߦ߅ߌࠆ⚵឵߃ߩ↪ߩߚ ߩᜰ㊎ߦޠၮߠ߈ޔᣣᧄߢߩᩱၭߦߟߡޔᜰ㊎߳ߩㆡวᕈ ߇⏕ߐࠇߚޕ 19 2004 ᐕ 6 ㄘᨋ᳓↥⋭߮ⅣႺ⋭ࠃࠅㆮવሶ⚵ߺ឵߃↢‛╬ߩ↪╬ߩ ⷙߦࠃࠆ↢‛ߩᄙ᭽ᕈߩ⏕ߦ㑐ߔࠆᴺᓞߦၮߠߊ╙৻⒳ ↪ⷙቯߩᛚࠍฃߌߚޕ㧔㘩↪߹ߚߪ㘺ᢱ↪ߦଏߔࠆߚߩ ↪ޔᩱၭޔടᎿޔ▤ޔㆇ៝߮ᑄ᫈ਗ߮ߦߎࠇࠄߦઃ㓐ߔ ࠆⴕὑߦߟߡ㧕 ޣMON 88017MON 810 ߩ⢒ᚑߩ⚻ㆊޤ ᧄࠬ࠲࠶ࠢ♽⛔࠻࠙ࡕࡠࠦࠪߪޔMON 88017 ߣ MON 810 ߩ⥄ᱺ♽⛔ࠍᓥ᧪ߩ 㔀⢒⒳ᴺࠍ↪ߡߒߚ(p20 ߩ࿑ 5)ޕ ␠ᄖ⒁ߦߟ߈㕖㐿␜ ࿑ 3 㒰⨲ࠣࡎࠨ࠻⠴ᕈ߮ࠦ࠙࠴ࡘ࠙⋡ኂ⯻ᛶ᛫ᕈ࠻࠙ࡕࡠࠦࠪ MON88017 ߩ⢒ᚑ࿑ ␠ᄖ⒁ߦߟ߈㕖㐿␜ ࿑ 4 ⚵឵߃࠻࠙ࡕࡠࠦࠪ MON 810 ߩ⢒ᚑ࿑ ␠ᄖ⒁ߦߟ߈㕖㐿␜ ࿑ 5 ࠬ࠲࠶ࠢ♽⛔࠻࠙ࡕࡠࠦࠪ MON 88017MON 810 ߩ⢒ᚑ࿑ 20 (4) ⚦⢩ౝߦ⒖ߒߚᩭ㉄ߩሽ⁁ᘒ߮ᒰᩭ㉄ߦࠃࠆᒻ⾰⊒ߩቯᕈ ޣMON 88017 ߦ⒖ߒߚᩭ㉄ߩሽ⁁ᘒ߅ࠃ߮ᒻ⾰⊒ߩቯᕈޤ MON 88017 ߩࠨࠩࡦࡉࡠ࠶࠻ಽᨆߦࠃࠆᝌㆮવሶߩ⸃ᨆߩ⚿ᨐ⚵ᧄޔ឵߃࠻ ࠙ࡕࡠࠦࠪߩࠥࡁࡓਛ 1 ࡩᚲߦ 1 ࠦࡇߩ T-DNA 㗔ၞ߇⚵ߺㄟ߹ࠇߡࠆߎߣ߇ ⏕ߐࠇߚ(MON 88017 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠᷝ⾗ᢱ 2 ߩ Appendix III ߩ p13 ߩ Figure 3)⸃ߩߎޕᨆ⚿ᨐ߮Ⴎၮ㈩ߩ⸃ᨆ⚿ᨐ߆ࠄࠄ߆ߦߥߞߚᝌㆮવ ሶߩᮨᑼ࿑ࠍ࿑ 6(p22)ߦ␜ߒߚߦᦝޕᝌㆮવሶߪቯߒߡᓟઍߦㆮવߒߡࠆ ߎߣ߇ⶄᢙઍ(↪ߚઍߦߟߡߪ p20 ߩ࿑ 3 ߩ*ශߩߟߚ♽⛔ࠍෳᾖ)ߦ߅ ߌࠆࠨࠩࡦࡉࡠ࠶࠻ಽᨆࠃߞߡ␜ߐࠇߚ(MON 88017 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠ ᷝ⾗ᢱ 3 ߩ p26 ߩ࿑ 3-4)ޔߚ߹ޕ㒰⨲ࠣࡎࠨ࠻⠴ᕈ߮ࠦ࠙࠴ࡘ࠙⋡ኂ⯻ ᛶ᛫ᕈ߽ⶄᢙઍߢቯߒߡ⊒ߒߡࠆߎߣࠍㆬᛮߩㆊ⒟ߢࠣࡎࠨ࠻ᢔᏓ ⹜㛎߮ Cry3Bb1 Ⱞ⊕⾰ߩ᛫ࠍ↪ߚ ELISA ᴺߦࠃࠅ⏕ߒߡࠆޕ ޣMON 810 ߦ⒖ߒߚᩭ㉄ߩሽ⁁ᘒ߅ࠃ߮ᒻ⾰⊒ߩቯᕈޤ MON810 ߩࠨࠩࡦࡉࡠ࠶࠻ಽᨆߦࠃࠆᝌㆮવሶߩ⸃ᨆߩ⚿ᨐޔMON810 ߩࠥ ࡁࡓߩ 1 ࡩᚲߦ 1 ࠦࡇߩ cry1Ab ㆮવሶ⊒ߦᔅⷐߥ PV-ZMBK07 ↱᧪ߩ DNA ᢿ ߇⚵ߺㄟ߹ࠇߡࠆߎߣ߇⏕ߐࠇߚ(MON810 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠߩ ᷝ⾗ᢱ 2 ߩ p21 ߩ࿑ 3)⸃ߩߎޕᨆ⚿ᨐ߮Ⴎၮ㈩ߩ⸃ᨆ⚿ᨐ߆ࠄࠄ߆ߦߥ ߞߚᝌㆮવሶߩᮨᑼ࿑ࠍ࿑ 7(p23)ߦ␜ߒߚޔߚ߹ޕᝌㆮવሶߪቯߒߡᓟઍ ߦㆮવߒߡࠆߎߣ߇ⶄᢙઍ(p22 ߩ࿑ 4 ߩฝ⢋ߦ d ශߩߟߚઍ)ߦ߅ߌࠆࠨ ࠩࡦࡉࡠ࠶࠻ಽᨆߦࠃߞߡ␜ߐࠇߚ(MON810 ߩ↢‛ᄙ᭽ᕈᓇ㗀⹏ଔᦠᷝ⾗ᢱ 2 ߩ p26 ߩ࿑ 6 ߮ᷝ⾗ᢱ 3 ߩ p37 ߩ࿑ 2)⋡࡚࠙࠴ޔߚ߹ޕኂ⯻߳ߩᛶ᛫ᕈ߽ ⶄᢙઍߢቯߒߡ⊒ߒߡࠆߎߣࠍ↢‛ᬌቯߦࠃࠅㆬᛮߩㆊ⒟ߢ⏕ߒߡ ࠆޕ ዏޔMON810 ߩࠨࠩࡦࡉࡠ࠶࠻ಽᨆߦࠃࠆᝌㆮવሶ⸃ᨆߩ⚿ᨐࠦࡠࡕ࠙࠻ޔ ࠪߩࠥࡁࡓਛߦᝌߐࠇߚߩߪ PV-ZMBK07 ↱᧪ߩ cry1Ab ㆮવሶ⊒ߦᔅⷐߥ 㗔ၞߩߺߢޔnptII ㆮવሶ߿ PV-ZMGT10 ↱᧪ߩᡷᄌဳ cp4 epsps ㆮવሶߣ gox ㆮવ ሶߩ⊒ࠞ࠶࠻ߪሽߒߥߎߣ߇⏕ߐࠇߚ (MON810 ߩ↢‛ᄙ᭽ᕈᓇ㗀 ⹏ଔᦠߩᷝ⾗ᢱ 2 ߩ p22 ߩ࿑ 4 ߮ p23 ߩ࿑ 5)ޕ 21 6 cp4 epsps ract1 intron P-e35S wt CAB leader ract1 intron cry3Bb1 22 ᧄ࿑ߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ NOS 3’ Xho I 7064 Pst I 7080 tahsp17 3’ RB Hind III 3612 Pst I 2798 Pst I 3622 Xba I 4265 Pst I 2576 CTP2 Xho I 317 Hind III 310 P-ract ࿑ 6 MON 88017 ߩᝌㆮવሶ࿑ 6 LB ࿑7 MON 810 ߩᝌㆮવሶ࿑ 7 ࿑ 7 MON 810 ߩᝌㆮવሶ࿑7 7 ᧄ࿑ߦ⸥タߐࠇߚᖱႎߦଥࠆᮭ߮ౝኈߩ⽿છߪᣣᧄࡕࡦࠨࡦ࠻ᩣᑼળ␠ߦᏫዻߔࠆޕ 23