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一分子ＤＮＡのＧＣコンテント分布計測における長さ補正

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(1)2006−BIO−6 （9）   2006／9／15. 社団法人 情報処理学会 研究報告 IPSJ SIG Technical Report. ৻ಽሶ㧰㧺㧭ߩ㧳㧯ࠦࡦ࠹ࡦ࠻ಽᏓ⸘᷹ߦ߅ߌࠆ㐳ߐ⵬ᱜ The length calibration on GC content distribution measurement for single molecular analysis of DNA ⮮ᧄ ⍮ਯ㧘᧖ᶆ ᔘ↵㧘㐳ደ ጪᔒ㧘૒⮮ ືᄢ㧘Ḋ ዊᄥ㇢ ᄹ⦟వ┵⑼ቇᛛⴚᄢቇ㒮ᄢቇ ᖱႎ⑼ቇ⎇ⓥ⑼ ᖱႎ↢๮⑼ቇኾ᡹ Tomoyuki FUJIMOTO㧘Tadao SUGIURA㧘Takeshi NAGAYA㧘Tetsuo SATO㧘Kotaro MINATO Department of Bioinformatics and Genomics㧘 Nara Institute of Science and Technology. ߎࠇ߹ߢߦᚒ‫ࠬ࡜ࠟߪޘ‬ၮ᧼਄ߦ㧰㧺㧭ࠍિᒛ࿕ቯߒߡ㧳㧯․⇣⊛ߦⰯశᨴ⦡ߒ㧘ࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ࠍ↪޿ ߡ㧰㧺㧭ࠍ৻ಽሶࠗࡔ࡯ࠫࡦࠣߔࠆߎߣߢ㧰㧺㧭ߩ㧳㧯ࠦࡦ࠹ࡦ࠻ߩ৻ᰴరಽᏓࠍ⸘᷹ߢ߈ߚ㧚ߒ߆ߒࠟ࡜ࠬၮ ᧼਄ߦ࿕ቯߐࠇߚ㧰㧺㧭ߩන૏㐳ߐᒰߚࠅߩႮၮᢙ߇⇣ߥࠆߚ߼㧘࠺࡯࠲ࡌ࡯ࠬߩ੍᷹ߣᔅߕߒ߽৻⥌ߒߥ޿㧚 ߘߎߢ㧘Ⴎၮᢙߩ㆑޿ࠍセᱜߔࠆᣂߚߥ⸃ᨆࡕ࠺࡞ߣߘߩㆡ↪⚿ᨐߦߟ޿ߡႎ๔ߔࠆ㧚. We have analyzed single molecular DNA with single molecular imaging by evanescent field microscope and identified the DNA by comparing measured results of one dimensional distribution of local GC content to estimation from genome database. The number of base pair within the unit length of the DNA fixed on the glass substrates are different from estimation, however, measured results have not matched with genome database well. We report on the new analytic-model and the new application result considering calibration of the difference of the base pair number.. 㧝㧚ߪߓ߼ߦ  ⃻࿷㧘ࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜╬ߩశቇᛛⴚߩㅴዷߦ઻޿㧘৻ಽሶࠍኻ⽎ߣߒߚ㜞ᗵᐲߥ⸘᷹߇ ⴕࠊࠇߡ޿ࠆ. 1)㧚శࠍ↪޿ߚ⸘᷹ߪ㕖⎕უ࡮㕖ធ⸅ߦኻ⽎‛ࠍⷰኤߢ߈ࠆߣ޿ߞߚ․㐳ࠍᜬߜ㧘. ∔∛ߩ⸻ᢿ࡮੍㒐࡮ᴦ≮ᴺߩ㐿⊒ߩߚ߼ߩࠥࡁࡓ⸃ᨆ⎇ⓥߦᔕ↪ߐࠇߡ޿ࠆ㧚ㄭᐕടㅦߔࠆߎߩ ࠥࡁࡓ⸃ᨆ⎇ⓥߢߪ㧘DNA ߩႮၮ㈩೉ᖱႎࠍ㜞ㅦߦหቯߔࠆߎߣ߇᳞߼ࠄࠇߡ޿ࠆ㧚Ⴎၮ㈩೉ߦ 㑐ߔࠆᖱႎࠍ㜞ㅦߦᓧࠆߎߣ߇ߢ߈ࠇ߫㧘ࠥࡁࡓ㈩೉࠺࡯࠲ࡌ࡯ࠬߣᲧセࠍⴕ߁ߎߣߦࠃࠅ࠙ࠖ ࡞ࠬߩ․ቯ߿੝⒳ߩ⊒⷗߇น⢻ߣߥࠅ㧘ᣧᦼߩᴦ≮ߦߟߥ߇ࠆߣᦼᓙߢ߈ࠆ㧚 ߘߎߢᚒ‫ ߪޘ‬DNA ߩዪᚲ⊛ߥ GC ࠦࡦ࠹ࡦ࠻ಽᏓߪ৻᭽ߢߪߥ޿ὐߦ⌕⋡ߒ㧘ࠛࡃࡀ࠶࠮ࡦ ࠻㗼ᓸ㏜ߢ DNA ࠍ৻ಽሶࠗࡔ࡯ࠫࡦࠣߔࠆߎߣߢ㧘Ⴎၮ㈩೉ᖱႎߩขᓧࠍ⹜ߺߚ 2)㧚ࠟ࡜ࠬၮ᧼ ਄ߦ DNA ࠍિᒛ࿕ቯߒ㧘GC ․⇣⊛ߥⰯశᨴ⦡ߣ DNA ৻᭽ߥⰯశᨴ⦡ࠍ⚵ߺวࠊߖ㧘ࠛࡃࡀ࠶ ࠮ࡦ࠻㗼ᓸ㏜ߢ৻ಽሶࠗࡔ࡯ࠫࡦࠣߒ㧘GC ࠦࡦ࠹ࡦ࠻ߩ৻ᰴరಽᏓᖱႎࠍ⸘᷹ߒߚ㧚ࠟ࡜ࠬၮ ᧼਄߳ߩ DNA િᒛ࿕ቯᣇᴺߪ㧘ࠬࡍ࡞ࡒࠫࡦ᳓ṁᶧߩỚᐲߦࠃࠅ DNA ߇ࠣࡠࡆࡘ࡯࡞⁁ᘒ߆ࠄ િᒛ⁁ᘒ߳ᄌൻߔࠆᕈ⾰ࠍ↪޿㧘ࡇࡍ࠶࠹ࠖࡦࠣߦࠃࠆ᳓ᵹࠍ೑↪ߒߚ. −63−. 3)㧚ߒ߆ߒ㧘ߎߩᣇᴺߢ.

(2) ߪિᒛᤨߦ DNA ߇᳓ᵹ߆ࠄฃߌࠆജߩᄌൻ㧘AT࡮GC ⚿วߦࠃࠆ⚿วജߩ㆑޿ߦ઻߁િᒛ᧦ઙ ߩᄌൻߦࠃࠅ㧘િᒛ࿕ቯߒߚ DNA ߩන૏㐳ߐᒰࠅߩႮၮᢙߦ஍ࠅ߇↢ߓ㧘ࠥࡁࡓ࠺࡯࠲ࡌ࡯ࠬ ਄ߩ੍᷹ߣᔅߕߒ߽৻⥌ߒߥ޿㧚 ࠺࡯࠲ࡌ࡯ࠬ਄ߩ੍᷹୯ߣ⸘᷹୯ߩ৻⥌ᐲࠍะ਄ߐߖࠆߚ߼ߦߪ㧘න૏㐳ߐᒰࠅߩႮၮᢙߩ㆑ ޿ࠍセᱜߔࠆᔅⷐ߇޽ࠆ㧚ߘߎߢ DNA ࠍ৻᭽ߦⰯశᨴ⦡ߒߡᓧࠄࠇࠆ↹௝ߩ↹⚛୯ᖱႎࠍ↪޿ ࠆߎߣߢ㧘න૏㐳ߐ޽ߚࠅߩႮၮᢙߩ㆑޿ࠍセᱜߔࠆᚻᴺࠍ⠨᩺ߒߚ㧚 ᧄⓂߢߪ GC ࠦࡦ࠹ࡦ࠻ߩ⸘᷹ᚻᴺߦߟ޿ߡㅀߴߚᓟ㧘න૏㐳ߐ޽ߚࠅߩ DNA ߩႮၮᢙߩセ ᱜᴺ෸߮⸘᷹⚿ᨐࠍ␜ߒ㧘ᓥ᧪ᴺߣߩᲧセߦߟ޿ߡႎ๔ߔࠆ㧚. 2. 1. 3. 4. 6. 5. 20Ǵm       (a) YOYO-1 Ⱟశ↹௝            (b) 7-AAD Ⱟశ↹௝ ࿑ 1㧚િᒛ࿕ቯߒߚ DNA ߩ᷹ⷰ⚿ᨐ 2㧚GC ࠦࡦ࠹ࡦ࠻⸘᷹ᚻᴺ 㩷 48,502bp ߩdzDNA ࠍ↪޿䋬Ⱟశಽሶ 7-AAD ߦࠃࠅ GC ․⇣⊛ߦⰯశᨴ⦡ߒߚᓟ㧘YOYO-1 ߦ ࠃࠅ DNA ৻᭽ߦⰯశᨴ⦡ߔࠆ 4)㧚ߎߩ 2 ㊀ᨴ⦡ߒߚ DNA ࠍࠟ࡜ࠬၮ᧼਄ߦિᒛߐߖ㧘ฦⰯశಽ ሶࠍࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ߦࠃࠅബ⿠ߐߖ᠟ᓇࠍⴕ߁㧚ߎࠇࠄⰯశಽሶߩബ⿠ᵄ㐳dzex ߣⰯశᵄ 㐳dzem ߪ㧘7-AAD ߢߪdzex=550 nm㧘dzem=650 nm㧘YOYO-1 ߢߪdzex=490 nm㧘dzem=510 nm ߢ޽ࠅ㧘ᵄ㐳ߩ㆑޿߆ࠄኈᤃߦ⼂೎น⢻ߢ޽ࠆ㧚ࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ߪ㧘ࠛࡃࡀ࠶࠮ࡦ࠻⁁ᘒ ߣߥߞߚశࠍബ⿠శߣߒߡ೑↪ߔࠆⰯశ㗼ᓸ㏜ߢ㧘ࡃ࠶ࠢࠣ࡜࠙ࡦ࠼ߩᓇ㗀ࠍฃߌߕߦၮ᧼⴫㕙 ㄭறߩߺࠍⷰኤߔࠆߎߣ߇ߢ߈ࠆ 5)㧚 ࿑ 1(a)ߦ YOYO-1 ࠍബ⿠ߐߖߚ↹௝㧘(b)ߦ 7-AAD ࠍബ⿠ߐߖࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ߦࠃࠅ᠟ ᓇߒߚ↹௝ࠍ␜ߔ㧚ฦ↹௝ߪ 12bit Ớ᷆↹௝ߢ㧘1 ㄝߪ 100Ǵm㧘1024 ↹⚛ߣߥߞߡ޿ࠆ㧚7-AAD ߩⰯశ↹௝ߪࠦࡦ࠻࡜ࠬ࠻߇ૐߊ DNA ߩ್೎⢻߇ഠࠆߚ߼㧘DNA ߩ᛽಴ߦߪ YOYO-1 Ⱟశ↹ ௝ࠍ↪޿ࠆ㧚YOYO-1 Ⱟశ↹௝ࠍ↪޿ DNA ✢ಽߩየᩮ✢ࠍ᛽಴ߒ↹⚛୯ fYOYO-1 ࠍ᳞߼ࠆ㧚የᩮ ✢ߩ᛽಴ߪ㧘DNA ߩਔ┵ 2 ὐࠍ⚿߱⋥✢ߦု⋥ߥⶄᢙߩ✢਄ߢᦨ߽㜞޿↹⚛୯ࠍ᳞߼ࠆߎߣߢⴕ. −64−.

(3) ߁㧚㓞ࠅว߁ု⋥ߥ✢ߩ㑆㓒ߪ↹௝਄ߢ᳞߼ߚ࡙࡯ࠢ࡝࠶࠼〒㔌ߩන૏㐳ߐߣߒߡ޿ࠆ㧚⛯޿ߡ 7-AAD Ⱟశ↹௝਄ߢ fYOYO-1 ߣหߓ૏⟎ߩ↹⚛୯ f7-AAD ࠍᓧࠆ㧚ߘߒߡ㧘(1)ᑼߦࠃࠅ GC ࠦࡦ࠹ ࡦ࠻ࠍ▚಴ߔࠆ 1)㧚ߎߩᠲ૞ࠍㆬᛯߒߚฦ DNA ߩࡇࠢ࠮࡞Ფߦⴕߞߡ޿ߊߎߣߢ㧘1 ᧄߩ DNA ߩ GC ࠦࡦ࠹ࡦ࠻ࠍ᳞߼ࠆߎߣ߇ߢ߈ࠆ㧚ߥ߅ DNA ㆬᛯᤨߦ߅ߌࠆየᩮ✢᛽಴ߢߪ GC ࠦࡦ࠹ ࡦ࠻ࠍ᳞߼ࠆߚ߼ߩ㑆㓒ߪ৻ቯߣߪ㒢ࠄߥ޿߇㧘ᧄႎ๔ߦ߅޿ߡߪߎߩࠨࡦࡊ࡝ࡦࠣ㑆㓒ࠍ৻ቯ ߣᛒ߁㧚. GCcontent =. f 7- AAD ‫ޓޓ‬ [%]‫ޓޓޓޓޓ‬ (1) f YOYO-1. ߹ߚ 7-AAD ߣ YOYO-1 ߩⰯశᒝᐲߩ㆑޿ߦߟ޿ߡߪ㧘dzDNA ߩ GC ࠦࡦ࠹ࡦ࠻ߩℂ⺰୯ߣ⸘ ᷹୯ߩᐔဋ୯߇৻⥌ߔࠆߣ઒ቯߒ⵬ᱜߒߡ޿ࠆ㧚 3㧚න૏޽ߚࠅ㐳ߐߩセᱜᴺ  ࿑ 2 ߦ࿑ 1(a)ߩ 1 ࠃࠅᓧߚ DNA ߩ↹⚛୯ಽᏓ㧘࿑ 3 ߦ(b)ࠃࠅᓧߚหߓ૏⟎ߦ޽ࠆ DNA ߩ↹ ⚛୯ಽᏓࠍ␜ߔ㧚Ⱟశಽሶ YOYO-1 ߪ DNA ࠍ৻᭽ߦᨴ⦡ߔࠆߚ߼㧘࿑ 2 ߩ↹⚛୯ಽᏓ߽৻᭽ߣ ߥߞߡ޿ࠆߴ߈ߢ޽ࠆ㧚ߒ߆ߒ DNA ߩિᒛᐲว޿߇৻ቯߢߥ޿ߎߣ߆ࠄ㧘↹⚛୯ಽᏓߦ஍ࠅ߇ ↢ߓߡ޿ࠆ㧚ߘߎߢ㧘ߎߩ↹⚛୯ಽᏓࠍၮߦ DNA ߩિ❗ࠍផቯߔࠆ㧚߹ߚ࿑ߢߪ DNA ਔ┵ߩ↹ ⚛୯߇ઁߩ႐ᚲߣᲧߴᭂ┵ߦ㜞ߊߥߞߡ޿ࠆ㧚ߎߩℂ↱ߣߒߡߪ㧘DNA ߇ࠣࡠࡆࡘ࡯࡞⁁ᘒߩ߹ ߹િᒛߖߕߦࠟ࡜ࠬၮ᧼਄ߦ࿕ቯߐࠇߡ޿ࠆ߽ߩߣ⠨߃ࠄࠇࠆ㧚ߘߩߚ߼િᒛࠍផቯߔࠆ೨ಣℂ ߣߒߡਔ┵ࠍኻ⽎㗔ၞᄖߣߒ㧘ਛᄩㇱಽߩᖱႎߩߺࠍ↪޿ߡන૏޽ߚࠅ㐳ߐߩセᱜࠍⴕ߁㧚   YOYO-1 ߩ↹⚛୯߇㜞޿႐ᚲߪ 1 ↹⚛ߦ฽߹ࠇࠆⰯశಽሶߩᢙ߇ᄙ޿㧘ߔߥࠊߜ฽߹ࠇߡ޿ࠆ Ⴎၮᢙ߽ᄙ޿ߣ⠨߃ࠄࠇ෼❗ߒߡ޿ࠆ߽ߩߣߔࠆ㧚ㅒߦ↹⚛୯ಽᏓߩૐ޿႐ᚲߪⰯశಽሶ߇ዋߥ ޿ߚ߼ DNA ಽሶᢙ߇ዋߥߊિᒛߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ㧚ߎߎߢߪિᒛࠍታ⃻ߔࠆߚ߼ߦએਅߩ 3 ⒳㘃ߩિᒛᴺߦߟ޿ߡᲧセᬌ⸛ࠍⴕߞߚ㧚. 㪋㪌㪇㪇 㪋㪇㪇㪇 㪊㪌㪇㪇 ↹⚛୯. 㪊㪇㪇㪇 㪉㪌㪇㪇 㪉㪇㪇㪇 㪈㪌㪇㪇 㪈㪇㪇㪇 㪌㪇㪇 㪇 㪇. 㪈. 㪉. 㪊. 㪋. 㪌. 㪍. 㪎 㪏 㪐 㪈㪇 㪈㪈 㪈㪉 㪈㪊 㪈㪋 㪈㪌 㪈㪍 㪧㫆㫊㫀㫋㫀㫆㫅㪲㱘㫄㪴. ࿑ 2㧚 YOYO-1 ߩ↹⚛୯ಽᏓ. −65−.

(4) ↹⚛୯. 㪈㪇㪇㪇 㪐㪇㪇 㪏㪇㪇 㪎㪇㪇 㪍㪇㪇 㪌㪇㪇 㪋㪇㪇 㪊㪇㪇 㪉㪇㪇 㪈㪇㪇 㪇 㪇. 㪈. 㪉. 㪊. 㪋. 㪌. 㪍. 㪎 㪏 㪐 㪈㪇 㪈㪈 㪈㪉 㪈㪊 㪈㪋 㪈㪌 㪈㪍 㪧㫆㫊㫀㫋㫀㫆㫅㪲㱘㫄㪴. ࿑ 3㧚  7-AAD ߩ↹⚛୯ಽᏓ 1). YOYO-1 ߩ↹⚛୯ߦኻߒ✢ᒻߦિᒛ  YOYO-1 ߩ↹⚛୯ߩᦨᄢ୯(max)ߣᦨዊ୯(min)ࠍ᳞߼㧘િᒛᐲว޿ࠍ↹⚛୯ߦኻߒ✢ᒻߦ ᄌൻߐߖࠆ㧚DNA ߇િᒛߒߡ޿ࠆߣߺߥߖࠆ↹⚛୯ߩૐ޿૏⟎ߪන૏㐳ߣߒ㧘↹⚛୯߇ᦨᄢ୯ ߦㄭߠߊߦߟࠇ㐳ߐࠍ n Ბ㓏ߢ✢ᒻߦᄌൻߐߖࠆ㧚޽ࠆ↹⚛୯(fYOYO-1)ࠍᜬߟὐߩિᒛᐲ(ex) ߪ(2)ᑼߢ⸘▚ߔࠆ㧚੹࿁ߪ n=10 ߣߒ⸘▚ࠍⴕߞߚ㧚. ex. fYOYO-1  min un max  min. ( 2). ߎߩߣ߈㧘7-AAD ߩ↹⚛୯ಽᏓߦኻߒߡ߽ YOYO-1 ߩ↹⚛୯ಽᏓࠍၮߦห᭽ߩિᒛᠲ૞ࠍⴕ ߁㧚߹ߚિᒛᓟߩ 7-AAD ߩ↹⚛୯(f ·7-AAD)ߪ(3)ᑼߩࠃ߁ߦિᒛᐲ߇Ⴧߔߏߣ 10%ߕߟ୯ࠍૐߊ ߔࠆࠃ߁ߦᠲ૞ࠍⴕߞߡ޿ࠆ㧚↹⚛୯߇ᦨᄢ୯ߥࠄ㐳ߐߪ 10 ୚㧘f ·7-AAD ߪ f7-AAD ߩ 10%ߩ୯ߣ ߥࠆ㧚. f7c- AAD. f7 - AAD  f7 - AAD u 0.1(ex  1). (3). 2). YOYO-1 ߩ↹⚛୯ߦኻߒᜰᢙ⊛ߦિᒛ  ਄ㅀߒߚᚻᴺ 1 ߣห᭽ߦ YOYO-1 ߩᦨᄢ୯ߣᦨዊ୯ࠍ૶↪ߒ㧘(4)ᑼߩࠃ߁ߦિᒛᐲࠍᜰᢙ⊛ ߦᄌൻߐߖߚ㧚. ex = ab n ߎߎߢ a = exp. ( 4). 㧙 min× ln(n ) ln(n ) 㧘b = ߢ᳞߼ߚᜰᢙᦛ✢ߩࡄ࡜ࡔ࡯࠲ߣߔࠆ㧚 max- min max 㧙 min. 3).YOYO-1 ߩන૏↹⚛୯Ფߦ 1 ߕߟિᒛ  YOYO-1 ߩ↹⚛୯߇න૏↹⚛୯ࠍ⿥߃ࠆᲤߦ 1 ߕߟિᒛߐߖࠆ㧚න૏↹⚛୯ࠍ 500 ߣߒ㧘↹ ⚛୯߇ 500 એਅߢߪන૏㐳㧘એ㒠↹⚛୯߇ 500 Ⴧ߃ࠆᲤߦන૏㐳ߩ 2 ୚㧘3 ୚ߣߒ㧘ᦨᄢ୯ߢ ޽ࠆ 4095 ߢߪ 9 ୚ߣߥࠅ㧘n=9 ߣߥࠆ㧚ߎߩᣇᴺߪ਄⸥ 2 ߟߩᦨᄢ୯ߣᦨዊ୯ࠍ↪޿㧘⋧ኻ. −66−.

(5) ⊛ߦિᒛߐߖࠆᚻᴺߣߪ⇣ߥࠅ㧘YOYO-1 ߩ↹⚛୯ಽᏓ߇⇣ߥߞߡ߽↹⚛୯߇หߓߢ޽ࠇ߫િ ᒛᐲว޿ߪ৻ቯߣߥࠆ㧚 ਄⸥ 3 ᚻᴺࠍ↪޿ DNA ߩන૏㐳ߐߩ㆑޿ࠍౣ⸘▚ߔࠆߎߣߢિᒛߐߖߚᓟ㧘੹࿁૶↪ߒߚ  dzDNA ߩℂ⺰਄ߩ㐳ߐߢ޽ࠆ ⚂ 16[Ǵm]߹ߢో㐳 DNA ࠍ✢ᒻߦ❗߼ࠆߎߣߢන૏޽ߚࠅ㐳ߐ ߩセᱜࠍⴕߞߡ޿ࠆ㧚࿑ 4 ߪdzDNA ߩ GC ࠦࡦ࠹ࡦ࠻ߩℂ⺰୯ߢ㧘ࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ߩಽ⸃ ⢻ࠍ⠨ᘦߒන૏㐳ߐ޽ߚࠅߩႮၮᢙࠍ 600bp ߣ઒ቯߒߡ޿ࠆ 2)㧚dzDNA ߩ GC ࠦࡦ࠹ࡦ࠻ߪ࿑ ਛ٤ߩࠃ߁ߦਛᄩઃㄭߦ․ᓽ⊛ߥ⪭ߜㄟߺ߇⷗ࠄࠇࠆ㧚ߎߩ․ᓽ⊛ߥ⪭ߜㄟߺㇱಽࠍ৻⥌ߐߖߚ ᓟߦో㐳 DNA ࠍ❗߼ߡ޿ࠆ㧚. 㪎㪇.  ᠟ᓇߒߚ DNA ߩ 6 ᧄߦߟ޿ߡ. 㪍㪇. ⵬ᱜߥߒ㧘෸߮િᒛࠍ⠨ᘦߒߚ߽ ߩߦߟ޿ߡᲧセࠍⴕߞߚ㧚6 ᧄߩ DNA ߩ߁ߜᦨ⍴ߪ⚂ 11Ǵm㧘ᦨ 㐳ߢ⚂ 25Ǵm㧘ᐔဋߪ⚂ 16Ǵm. 㪞㪚㪺㫆㫅㫋㪼㫅㫋㪲㩼㪴. 4㧚⸘᷹⚿ᨐ. 㪌㪇 㪋㪇 㪊㪇 㪉㪇 㪈㪇 㪇. ߣߥߞߚ㧚. 㪇. 㪊. 㪍. 㪐 㪈㪉 㪈㪌 㪈㪏 㪉㪈 㪉㪋 㪉㪎 㪊㪇 㪊㪊 㪊㪍 㪊㪐 㪋㪉 㪋㪌 㪋㪏 ૏⟎㪲㫂㪹㫇㪴. ℂ⺰୯ߣ⸘᷹୯ߩ㘃ૃᐲࠍ್. ࿑ 4㧚dzDNA ߩ GC ࠦࡦ࠹ࡦ࠻. ᢿߔࠆߚ߼ߦ(5)ᑼߩ⋧㑐ଥᢙࠍ ᳞߼ߚ㧚 N. C. 㺌^ DB i

(6)  DB

(7) M i

(8)  M

(9) ` GC. GC. GC. GC. i 1. N. N. 㺌^DB i

(10)  DB ` 㺌^M i

(11)  M ` GC. i 1. GC. 2. GC. GC. 2. ‫ޓޓޓޓޓޓ‬ (5). i 1. ߎߎߢ㧘DBGC(i)ߪ i ⇟⋡ߩℂ⺰୯ߩ GC ࠦࡦ࠹ࡦ࠻㧘 DBGC ߪℂ⺰୯ߩ GC ࠦࡦ࠹ࡦ࠻ߩᐔဋ ୯㧘MGC(i)ߪ i ⇟⋡ߩ⸘᷹୯ߩ GC ࠦࡦ࠹ࡦ࠻㧘 M GC ߪߘߩᐔဋ୯ߣߥࠆ㧚߹ߚ N ߪℂ⺰୯ߩ ࠺࡯࠲ᢙߣߥࠆ㧚(5)ᑼࠍ↪޿ߡ࿑ 1(a)ߩ DNA㧘1 ߆ࠄ 6 ⇟ߩᲧセࠍⴕߞߚ⚿ᨐࠍ⴫ 1 ߦ␜ߔ㧚 ߹ߚ㧘DNA1 ߩ GC ࠦࡦ࠹ࡦ࠻ࠍ࿑ 5 ߦ㧘DNA6 ߩ GC ࠦࡦ࠹ࡦ࠻ࠍ࿑ 6 ߦ㧘(a)ߪセᱜߥߒ㧘(b) ߪᚻᴺ 1 ࠍㆡ↪ߒߚ߽ߩࠍ␜ߔ㧚 ⴫ 1㧚GC ࠦࡦ࠹ࡦ࠻ߩℂ⺰୯ߣ⸘᷹୯ߩᲧセ. &0#      . ᚻᴺ      . ⋧㑐ଥᢙ ᚻᴺ ᚻᴺ            . セᱜߥߒ      . −67−. ⴫ 2㧚ਛᄩㇱߩߺߩᲧセ. &0#      . ᚻᴺ      . ⋧㑐ଥᢙ ᚻᴺ ᚻᴺ            . セᱜߥߒ      .

(12)  . )%EQPVGPV=?.    . Database Measured.    . . . . . . .    ૏⟎=MDR?. . . . . . . . (a) ℂ⺰୯ߣセᱜߥߒ࠺࡯࠲ߩᲧセ  . )%EQPVGPV=?.    . Database Measured.    . . . . . . . . . . . . . . ૏⟎=MDR?. (b) ℂ⺰୯ߣᚻᴺ 1 ߩᲧセ ࿑ 5㧚 ࿑ 1(a)ਛ 1 ߩ GC ࠦࡦ࠹ࡦ࠻ߩℂ⺰୯ߣ⸘᷹୯ߩᲧセ. −68−. . . .

(13)  . )%EQPVGPV=?.   . Database Measured.    . . . . . . .    ૏⟎=MDR?. . . . . . . . . . . . . (a) ℂ⺰୯ߣセᱜߥߒ࠺࡯࠲ߩᲧセ.  . )%EQPVGPV=?.       . . . . . . .    ૏⟎=MDR?. . . (b) ℂ⺰୯ߣᚻᴺ 1 ߩᲧセ ࿑ 6㧚 ࿑ 1(a)ਛ 6 ߩ GC ࠦࡦ࠹ࡦ࠻ߩℂ⺰୯ߣ⸘᷹୯ߩᲧセ. −69−.

(14) ⴫ 1 ࠃࠅ㧘િᒛࠍ⠨ᘦߒߚ႐วߦଥᢙߩะ਄߇⷗ࠄࠇߚߩߪ 6 ᧄਛ 5 ᧄߣߥߞߚ㧚࿑ 1(a)ਛ 1 ߦ㒢ࠅセᱜߒߥ޿႐วߩଥᢙ߇㜞߆ߞߚ㧚߹ߚ㧘࿑ 1(a)ਛ 6 ߪિᒛࠍ⠨ᘦߔࠆߎߣߢ⋧㑐ଥᢙ߇ ะ਄ߒߡ޿ࠆ㧚ߒ߆ߒ㧘ో૕⊛ߦଥᢙߪ㜞޿ߣߪ޿߃ߥ޿㧚ේ࿃ߣߒߡ㧘ਛᄩㇱߩ⪭ߜㄟߺߩ޽ ࠆ㗔ၞߪ৻⥌ߒߡ޿ࠆ߇㧘ਔ┵ߢߩ⚦߆ߥᏅ⇣߇⠨߃ࠄࠇࠆ㧚ߘߎߢ㧘ਛᄩㇱ 12kbp ߆ࠄ 36kbp ઃㄭߦ⌕⋡ߒ㧘ౣ⸘▚ߒߚ⋧㑐ଥᢙࠍ⴫ 2 ߦ␜ߔ㧚 ⴫ 2 ߣ⴫ 1 ࠍᲧߴࠆߣඨᢙએ਄ߩଥᢙߢ୯ߩะ਄߇⏕⹺ߢ߈ߚ㧚․ߦ࿑ 1(a)ਛߩ 1 ߣ 2 ߦߟ޿ ߡߪోߡߦ߅޿ߡะ਄߇⷗ࠄࠇߚ㧚ߒ߆ߒ࿑ 1(a)ਛ 3 ߦߟ޿ߡߪセᱜߒߚ୯ోߡ߇⴫ 1 ࠃࠅᷫዋ ߒߡ޿ࠆ㧚߹ߚ㧘ᚻᴺߏߣߦ⷗ࠆߣ 1 ߢߪ⴫ 1 ߦᲧߴో૕⊛ߦଥᢙ߇ะ਄ߒߡ޿ࠆ߇ᚻᴺ 2 ෸߮ 3 ߢߪ౒ߦඨᢙߩะ਄ߦߣߤ߹ߞߚ㧚ߎߎߢߪ㧘ᜰᢙ⊛ߦિᒛߐߖࠆᚻᴺ 2 ࠍㆡ↪ߒߚ⚿ᨐࠃࠅ ߽✢ᒻߦિᒛߐߖࠆᚻᴺߢ޽ࠆ 1㧘3 ࠍㆡ↪ߔࠆᣇ߇㜞޿ଥᢙࠍ␜ߒߚ㧚 5㧚߹ߣ߼  DNA િᒛ࿕ቯᤨߦ↢ߓࠆන૏㐳ߐ޽ߚࠅߩႮၮᢙߩ㆑޿ࠍセᱜߔࠆߚ߼ߦ㧘৻ಽሶ DNA ߩ GC ࠦࡦ࠹ࡦ࠻⸘᷹ᤨߦ߅ߌࠆિᒛࠍ⠨ᘦߒ㧘න૏޽ߚࠅ㐳ߐࠍセᱜߒߚ⚿ᨐߦߟ޿ߡㅀߴߚ㧚 િᒛࠍ⠨ᘦߒන૏޽ߚࠅ㐳ߐࠍセᱜߔࠆߎߣߢ࠺࡯࠲ࡌ࡯ࠬߣߩ㘃ૃᐲࠍะ਄ߐߖࠆߎߣ߇ߢ߈ ߚ㧚߹ߚ㧘િᒛᣇᴺߦߟ޿ߡߪ㧘ᜰᢙ⊛ߦિᒛߐߖࠆࠃࠅ߽✢ᒻ⊛ߦિᒛߐߖࠆ߶߁߇ࠃࠅ㘃ૃ ᐲ߇ะ਄ߒߚ㧚ᧄႎ๔ߢߪિᒛߐߖߚ࠺࡯࠲ࠍ෼❗ߐߖࠆ㓙ߦ✢ᒻᚻᴺࠍ↪޿ߚ߇㧘ታ㓙ߩ DNA ߩિᒛߪ㕖✢ᒻߢ޽ࠆߣᕁࠊࠇࠆߚ߼㧘GC ࠦࡦ࠹ࡦ࠻ࠍၮߦฦಽሶߩ⚿วജߩ㆑޿ࠍ⠨ᘦߒߚ િᒛᴺߥߤࠍᬌ⸛ߔࠆᔅⷐ߇޽ࠆߣᕁࠊࠇࠆ 6)㧚߹ߚ㧘ࠛࡃࡀ࠶࠮ࡦ࠻㗼ᓸ㏜ߦࠃࠆᾖ᣿႐߇৻ ᭽ߢߪߥ޿ὐ߿㧘৻ቯߣ઒ቯߒߡ޿ࠆየᩮ✢ߩࠨࡦࡊ࡝ࡦࠣ㑆㓒ߩ⵬ᱜࠍⴕ߁ߎߣߢ㧘ߐࠄߥࠆ GC ࠦࡦ࠹ࡦ࠻ߩ㘃ૃᐲߩะ਄߇ᦼᓙߢ߈ࠆ㧚 ෳ⠨ᢥ₂ 1) Y.Harada 㧘 T.Funatsu 㧘 K.Murakami 㧘 Y.Nonoyama 㧘 A.Ishihara 㧘 and T.Yanagida 㧘 ̌Single-molecule imaging of RNA polymerase-DNA interactions in real time̍㧘Biophysical Journal Vol.76 February 1999㧘pp709-715 (1999)㧚 2) ᧖ᶆᔘ↵㧘㐳ደጪᔒ㧘૒⮮ືᄢ㧘Ḋዊᄥ㇢㧘̌GC ࠦࡦ࠹ࡦ࠻ߩ৻ᰴరಽᏓᖱႎࠍ↪޿ߚ৻ಽ ሶ DNA ߩ⸃ᨆ̍㧘 ᖱႎಣℂቇળ⎇ⓥႎ๔㧘 2006-BIO-004㧘 Vol㧚2006㧘No㧚13㧘pp25-32 (2006)㧚 3) Smith㧘S.㧘Y.Cui㧘and C.Bustamante㧘 ̌Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads̍㧘Science㧘Vol㧚258㧘pp1122-1126 (1992)㧚 4) R.Zucker㧘A.Keshaviah㧘O.Price㧘and J.Goldman㧘 ̌Confocal laser scanning microscopy of Rat Follicle development̍㧘The ournal of Histchemistry & Cytochemistry Vol㧚48(6)㧘 pp781-791 (2000)㧚 5) D.Axellrod㧘 ̌Total internal reflection fluorescence microscopy in cell biology̍㧘Traffic㧘Vol.2㧘 pp㧚764-774 (2001)㧚 ̌Estimating 6) C.Bouchiat㧘M.D.Wang㧘J.-F.Allemand㧘T.Strick㧘S.M.Block㧘and V.roquette㧘 the persistence length of a worm-like chain molecule from force-extension measurements̍㧘 Biophysical Journal Vol.76㧘January 1999㧘pp409-413 (1999)㧚. −70−.

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