次世代シーケンサーによるメタゲノム解析：桜の花びらに付着した環境DNAを解析する

DNA (DBCLS) 2018/12/26 JST

ࠓ೔ͷ͓࿩ ੜ෺ֶNFBUT৘ใՊֶ

http://docman.dbcls.jp/im/

ٕज़ͷਐา͕ή ϊϜՊֶͷઓུΛม͑ͨ ೥͝Ζ

Ή͔͠αϯΨʔ๏ʹΑΔ%/"഑ྻղಡ Ͱ͖Δ͜ͱ͔Βͭͭ͜͜ͱ

͜ΕΛઃܭ͠ͳ͍ͱಡΊͳ͍ ग़య: https://www.researchgate.net/ﬁgure/Principle‐of‐Sanger‐sequencing‐Incorporation‐of‐ ddNTPs‐terminates‐the‐elongation‐of‐the_ﬁg4_265264877

͍·େن໛Ԙج഑ྻղಡٕज़ʹΑΔ໢ཏతܭଌ ͱΓ͋͑ͣશ෦ಡΉ

Ϳͭ੾Γʹ͢Δ શ෦ʹΞμϓλʔ഑ྻΛ͚ͭΔ શͯͷ഑ྻʹରͯ͠ ૬ิ࠯ͷ߹੒Λ͢Δ ೋ࣍ݩ্ۭؒͷܬޫΛ ΧϝϥͰࡱӨɺܦ࣌ͷ มԽΛॲཧͯ͠഑ྻʹ ม׵͢Δ ग़య: https://bitesizebio.com/13546/sequencing‐by‐synthesis‐explaining‐the‐illumina‐sequencing‐technology/

ಡΊͳ͔ͬͨ΋ͷ͕ಡΊΔΑ͏ʹ ͱΓ͋͑ͣಡΜͰΈΑʂ

ग़య: https://gold.jgi.doe.gov/statistics

͋ͷੜ෺ͷή ϊϜͬͯ΋͏ಡ·Ε͚ͯͨͬʁ ή ϊϜห౰ϓϩδΣΫτ

ग़య: https://www.instagram.com/p/BcybbydAbSK/

ग़య: http://wiki.lifesciencedb.jp/mw/GenomeBentoProject

େྔฒྻΛ࢖༷ͬͯʑͳݱ৅Λܭଌ͢Δ 4FRͷ࣌୅

ϥϘ͔Βσʔλ͕ͲΜͲΜग़ͯ͘Δ ͕ͩγʔέϯα͸ੜମ෼ࢠΛจࣈʹม׵͢ΔػցͰ͋Γ஌ࣝΛੜΉ΋ͷͰ͸ͳ͍

ੜ෺ֶͷ஌ࣝ ৘ใՊֶͷٕज़Ͱ จࣈྻͷฒͼΛ஌ࣝʹม׵͢Δ

#JH%BUB4DJFODFͱͳͬͨੜ෺ֶ͸ܭࢉػͳ͠Ͱ͸΍͍͚ͬͯͳ͍ ੜ෺ֶNFBUT৘ใՊֶ

Պֶతख๏ͷͭͷύϥμΠϜ ܦݧཧ࿦ܭࢉσʔλ

Ծઆ͔Β࣮ݧΛσβΠϯ͢ΔPS·ͣσʔλΛऔͬͯ෼ੳ͢Δ Ծઆۦಈܕσʔλۦಈܕ

ϝλή ϊϜ Կ͕ೖͬͯΔ͔Θ͔Μͳ͍͚Ͳ%/"શ෦ಡΊ͹Α͘Ͷʁ

ւਫதͷ%/"͔ΒڕͷछྨΛਪఆ ஑ͷਫશ෦ಡΉ ग़య: https://doi.org/10.1098/rsos.150088-9

ۭؾதͷ%/"͔Βඍੜ෺Λਪఆ ۭؾಡΉ ग़య: https://doi.org/10.1371/journal.pone.0001862

ެڞۭؒͷ%/"͔Βඍੜ෺Λਪఆ .FUB46#1SPKFDU ग़య: https://doi.org/10.1186/s40168‐016‐0168‐z

ഉᔔ෺தͷ%/"͔Β௎಺ࡉەΛਪఆ ʜ͕Α͘ߦΘΕΔ͕Լͷਤ͸໺ੜಈ෺Λই͚ͭͣʹ%/"Λ࠾औ͢Δख๏ͷ࿦จ ग़య: https://doi.org/10.1038/s41598-018-20427-9

೥ౙɾ౎಺๭ॴ ݚڀूձʮୈ࢛ճ/(4ݱ৔ͷձʯاըձٞ

ʮ͜Ε͚ͩγʔέϯε҆͘ͳͬͨΜ͔ͩΒͳΜ͔ಡ·ͳ͍ʁʯ

ࡩͷ඼छͰ͋ΔιϝΠϤγϊ͸઀͗໦Ͱ૿͑ΔΫϩʔϯ Ұఆͷؾ৅৚݅ͰҰ੪։Ֆ͢Δ͕Ұि͔ؒΒे೔Ͱࢄͬͯ͠·͏

॓ओͷή ϊϜ͸ಉ͡Ͱ؀ڥ৚͚͕݅ͩҟͳΔ ෇ணͨ͠%/"ͷҧ͍͸؀ڥͷҧ͍Λ൓ө͢Δ͔ʁ

ͱ͸͍͑ҋӢʹγʔέϯε͢Ε͹Α͍ͱ͍͏Θ͚Ͱ͸ͳ͍ͷͰ ݚڀσβΠϯ Λ͠·͢

༧ࢉ૊Έ αϯϓϦϯάΩοτͱࢼༀͷௐୡ αϯϓϦϯάΛ͢Δ αϯϓϧ͔Β%/"Λநग़͢Δ 4S3/"ྖҬΛ૿෯ͯ͠γʔέϯε͢Δ ੜ෺෼ྨσʔλϕʔεʹ໰͍߹ΘͤͯಡΜͩ഑ྻ͔Βੜ෺छΛਪఆ͢Δ ֶձͰղੳίϯϖςΟγϣϯΛ΍Δ ࿦จΛॻ͘

Ϋϥ΢υιʔγϯά શࠃ͔ΒࢀՃऀΛืͬͯαϯϓϦϯάΛ͓ئ͍͢Δ

༧ࢉ૊Έ αϯϓϦϯάΩοτͱࢼༀͷௐୡ ΢ΣϒΞϓϦέʔγϣϯͷ࣮૷ͱެ։ એ఻ͱࢀՃऀืू ΩοτΛࢀՃऀʹૹΔ αϯϓϦϯάͯ͠΋Β͏ αϯϓϧΛૹͬͯ΋Β͏ αϯϓϧ৘ใΛॻ͍ͯొ࿥ͯ͠΋Β͏ ૹΒΕ͖ͯͨαϯϓϧ͔Β%/"Λநग़͢Δ 4S3/"ྖҬΛ૿෯ͯ͠γʔέϯε͢Δ
ੜ෺෼ྨσʔλϕʔεʹ໰͍߹ΘͤͯಡΜͩ഑ྻ͔Βੜ෺छΛਪఆ͢Δ ֶձͰղੳίϯϖςΟγϣϯΛ΍Δ ࿦จΛॻ͘

αϯϓϦϯάࢀՃऀ 149 ໊

ू·ͬͨαϯϓϧ਺ 594

શࠃ੍೼ (ԭೄΛআ͘)

ಡΜͰΈͨ

ಘΒΕͨ഑ྻͷ΄ͱΜͲ͕ ༿྘ମͱϛτίϯυϦΞ ίϯλϛͩ

ίϯλϛ $POUBNJOBUJPOԚછ Պֶ࣮ݧҰൠʹ͓͍ͯҙਤ͠ͳ͍෺࣭͕ࠞೖ͢Δ͜ͱ

ɾТɾʆ Ұൃউෛͩͱ͜͏͍͏͜ͱ΋͋Δ

αϯϓϦϯά࣌ʹՖͼΒͷද໘Λ ࡲΓ͗ͯ͢ࡩͷࡉ๔͕ࠞೖͨ͠ͱ ࢥΘΕΔͷਤ

࡞ઓมߋ ݟ͔ͭͬͨ഑ྻͷதʹԿ͔໘ന͍΋ͷ͸ͳ͍͔ʁ

΄ͱΜͲ༿྘ମͱϛτίϯυϦΞ·ΈΕ ΄΅ίϯλϛͱ૝૾͞ΕΔ͕ɺશ෦͕ͦ͏ͳͷ͔ʁ

ҧ͏২෺ʹ༝དྷ͢ΔͱࢥΘΕΔ഑ྻ γʔέϯεͷϛεͷՄೳੑ΋͋Δ͕ɺϛεϚονͷ਺Λߟ͑Δͱʜʁ

Cryptomeria japonica Ֆ࣌ӍࡩࢄΓΏͦ͘ͷલʹ

ɾТɾʆ ͜͏ͯ͜͠ͷੈքʹ·ͨҰͭ৽ͨͳτϦϏΞ͕ੜ·Εͨʜ

ر๬͸ͳ͍ͷ͔ ͜ͷ··Ͱ͸কདྷʹͭͳ͕Βͳ͍

ܚጯٛक़େֶઌ୺ੜ໋Պֶݚڀॴ ߥ઒άϧʔϓ ߥ઒ Տ໺ ౻ݪ ٢ా ߴҪ Ѩ෦ ࠤ౻ণ௚͞Μ

ٕज़һ ٕज़һ ֶੜ ॿڭ ॿڭ ॿڭ ।ڭत ൿॻ 0 50
100 150 200 count abe Arakawa Fujiwara Ishii Kono sato Takai Yoshida member * * εʔύʔςΫχγϟϯҰਓউͪ ݸਓຖͷαϯϓϦϯάʹ͓͚ΔόΫςϦΞ഑ྻ਺ ߥ઒࿨੖͞Μ ܚጯٛक़େ

0 1000 2000 3000 count abe Arakawa Fujiwara Ishii Kono
sato Takai Yoshida member 0 50 100 150 200 count abe Arakawa Fujiwara Ishii Kono sato Takai Yoshida member ٕज़һ ٕज़һ ֶ ॿڭ ॿ ॿڭ ।ڭत ൿॻ ٕज़һ ٕज़һ ֶ ॿڭ ॿ ॿڭ ।ڭत ൿॻ ༿྘ମɾϛτίϯυϦΞআ֎ ༿྘ମɾϛτίϯυϦΞؚΉ ՖคΛଟ͘ͱͬͯ͠·͍ͬͯΔ όΫςϦΞΛ༏ઌతʹͱΕ͍ͯΔਓ͕͍Δ 10 15 20 25 30 35 40 45 50 55 4 500 1000 1500 2000 2500 17 20 40 60 80 100 4 0 500 1000 1500 2000 2500 3000 17 0 10 20 30 40 50 4 0 250 500 750 1000 1250 1500 17 0 10 20 30 40 50 4 0 250 500 750 1000 1250 1500 17 0 5 10 15 20 25 30 4 200 400 600 800 1000 1200 1400 17 0 5 10 15 20 4 0 200 400 600 800 1000 1200 17 0 50 100 150 200 4 1500 1750 2000 2250 2500 2750 3000 17 0 20 40 60 80 100 120 4 0 500 1000 1500 2000 17 ٕज़һ ٕज़һ ֶੜ ॿڭ ॿڭ ॿڭ ।ڭत ൿॻ ༿྘ମɾϛτίϯυϦΞͷ഑ྻ਺ͱόΫςϦΞͷ഑ྻ਺ͷ૬ؔ ߥ઒࿨੖͞Μ ܚጯٛक़େ

഑ྻσʔλɺϝλσʔλɺ࿦จͦͷଞશ෦͜͜ʹ͋Γ·͢ EBUBECDMTKQdJOVUBOPPNHTFR

αϯϓϦϯάͷख๏Λ޻෉͢Ε͹ΑΓΑ͍σʔλ͕ಘΒΕΔ ͔΋ SF͓ՖݟϓϩδΣΫτ ٻΉϓϩδΣΫτϦʔμʔ

Պֶݚڀͱͯ͠੒ཱͤ͞Δ͜ͱ͚ͩͰ͸ͳ͍ ͓ՖݟϓϩδΣΫτਅͷ໨త

༧ࢉ૊Έ αϯϓϦϯάΩοτͱࢼༀͷௐୡ ΢ΣϒΞϓϦέʔγϣϯͷ࣮૷ͱެ։ એ఻ͱࢀՃऀืू ΩοτΛࢀՃऀʹૹΔ αϯϓϦϯάͯ͠΋Β͏ αϯϓϧΛૹͬͯ΋Β͏ αϯϓϧ৘ใΛॻ͍ͯొ࿥ͯ͠΋Β͏ ૹΒΕ͖ͯͨαϯϓϧ͔Β%/"Λநग़͢Δ 4S3/"ྖҬΛ૿෯ͯ͠γʔέϯε͢Δ
ੜ෺෼ྨσʔλϕʔεʹ໰͍߹ΘͤͯಡΜͩ഑ྻ͔Βੜ෺छΛਪఆ͢Δ ֶձͰղੳίϯϖςΟγϣϯΛ΍Δ ࿦จΛॻ͘

೔࣌΢ΣϒΞϓϦέʔγϣϯ͕ࣗಈೖྗ αϯϓϦϯάͷ৔ॴ Ң౓ܦ౓ߴ౓ (14Ͱࣗಈೖྗ ఱީ೔࣌ͱ(14৘ใΛݩʹؾ৅ி%#͔Βநग़ͯࣗ͠ಈิ׬ ։Ֆޙͷ೔਺೔࣌ͱ(14৘ใ͔Βಉ͘͡ࡩ։Ֆ଎ใΑΓࣗಈิ׬ ͦͷଞͷ৘ใࣗ༝هड़

ϓϩͷݚڀैࣄऀ͕ಉ͡ର৅ʹ͍ͭͯௐ΂Δ ؍࡯ͱهड़ͷ࣭ͱྔ

ඈௗࢁଆ ࢪઃೖΓޱ ࢪઃෑ஍಺ JRԦࢠӺଆ ύʔΫαΠυ λϫʔαΠυ ιϝΠϤγϊ ಓ࿏Ԋ͍ೖΓޱ ߴ౓80.5m αΫϥͷՖ1ݸ෼
ιϝΠϤγϊݩ໦ $BTFT4JNQMFUFSNTPOMZ ղऍ͸೉͍͕͠ɺػց͕ಡΊΔ

000353(ը૾000353_1)ʹ͍ͭͯɻ000361ͱͷ૬ҧ఺:ڳߴ௚ܘ56cmɻथߴ10.02m(ը૾000353_2தஈ)ɻथྸ͓Α ͼ২࠿ޙͷ೥਺͕ߴ͍ͱਪఆ͞Ε·͢ɻը૾ڳߴ෦ҎԼͷथൽ্ͷணੜ২෺౳ͷඃ෴཰80%௒ɻணੜ২෺౳͸ϋ ϦΨωΰέՊͳͲ᥌ଽྨ,ϊΩγϊϒͳͲ༽ࣃྨ,΢ϝϊΩΰέ΍ϨϓϥΰέͳͲ஍ҥମ͕ओͰɺΩϊίྨ͸ݟ౰ͨ Γ·ͤΜͰͨ͠(ը૾000353_3)ɻৗ྘޿༿थΛओͱ͢Δೋ࣍ྛ·ͨ͸εεΩ૲஍͔Βͷڑ཭͸21mɻ000353ͱ 000361ͷڞ௨఺:ՖͷܗଶͷΈ͔ΒιϝΠϤγϊͱ൑அ͠·͕ͨ͠,౰஍Ͱ͸ެԂͳͲʹ͍ΘΏΔατβΫϥͷෳ਺ ඼छΛࠞ২͢Δ͜ͱ͕͋ΔͷͰ,ՄೳͰ͋Ε͹ը૾(000353_4)͔Β඼छͷ֬ೝΛ͓ئ͍͠·͢ɻࢼྉ࠾औ஍͸ࣛࣇ ౡࢢ಺ɻḈຎ൒ౡ౦ଆͷࣛࣇౡ࿷ʹ໘͠ɺՐࡅྲྀ෾ग़෺͕ଯੵͯ͠Ͱ্͖ͨϊݪ୆஍্ɻඪߴ90mɻ1978೥ʹ଄ ੒͞Εͨॅ୐஍֎ԑͷެԂʹ২࠿͞Εͨݸମɻ২࠿஍ͷެԂͷ๺౦ଆͱ๺੢ଆʹंಓΛִͯͯ,֓Ͷೋ֊ՈҰށݐ ͯͷॅ୐஍͕͋Γ·͢ɻ૬ରর౓100%ɻंಓ͓ΑͼंಓԊ͍ͷ֗౮͔Βͷڑ཭͸΄΅ಉ͡ɻࣛࣇౡ஍ํؾ৅୆෇ ۙͷԹྔࢦ਺͸140લޙɻௐࠪ஍͸ඪߴ͕80mఔ౓ߴ͍ͷͰͦΕΑΓΘ͔ͣʹ௿͍ͱߟ͑ΒΕ·͢ɻஆԹଳͷর༿
थྛଳʹ૬౰͠·͕͢,ஆԹଳͷর༿थྛଳʹ૬౰͠·͕͢,২࠿஍ͷެԂͷೆ੢ଆͷࣼ౓30°ͷ֑஍ͷࣼ໘ʹৗ྘ ޿༿थΛओͱ͢Δೋ࣍ྛ·ͨ͸૲஍͕࢒ଘ͍ͯ͠Δ͚ͩͰ͢ɻ࿈೔׆ൃʹ෾Ր͍ͯ͠Δࡩౡত࿨Րޱ͔Βͷڑ཭͸ ͓Αͦ15kmɻ෩޲͖ʹΑͬͯ͸߱փ͕͋Γ·͢ɻҰൠʹࣛࣇౡ࿷पลͰ͸ۭத࣪౓ͷߴ͞ͱՐࢁփͷ෇ணʹΑΓ थ໦ͷװʹணੜ২෺͕ੜҭ͠΍͍͢܏޲͕͋Γ·͢ɻαϯϓϧͷՖห্ʹ΋Րࢁփ͋Δ͍͸ԫ࠭ͱࢥΘΕΔԚΕ͕ ΈΒΕ·͢ɻҰൠతʹ,Ұ੪ʹ࡙͍ͯҰ੪ʹࢄΔؔ౦ͱൺ΂Δͱ,౰஍Ͱ͸1ຊͷथͷ։Ֆظ͕ؒ௕͍(ݸʑͷՖ͕ॱ ࡙͍࣍ͯ,ॱ࣍ࢄΔ)܏޲͕͋ΔΑ͏Ͱ͢ɻ্هͷ։Ֆޙͷ೔਺͸αϯϓϦϯάͨ͠ʮՖʯͷ։Ֆ೔਺Ͱ͢ɻҢ౓ܦ ౓ʹ͍ͭͯ͸GPSઐ༻ػͷ৘ใ(ը૾000353_2Լஈ)Λೖྗ͠·ͨ͠ɻ $BTFJOEFUBJM ྔ͸ଟ͍͕ɺػցՄಡੑ͸௿͍

هड़ͷίετWTղੳ࣌ͷॊೈੑ ϝλσʔλͷ࣭ͷ޲্͕EBUBESJWFOTDJFODFͷ伴ΛѲ͍ͬͯΔ

• ܭଌٕज़ͷਐา͕Պֶͷख๏Λม͑Δ • σʔλۦಈܕՊֶͱͳͬͨੜ෺ֶ͸ܭࢉػՊֶͱ༥߹͢Δ • ஝ੵͨ͠σʔλ͕ҖྗΛൃش͢Δ࣌୅ͦ͜σʔλͷهड़͕ॏཁ • ͍͔ʹ࣭ͷߴ͍σʔλΛऩू͢Δ͔ %

Պֶͱ͸هड़͢Δ͜ͱ ॻ͖ͱͲΊΑɻٞ࿦ͨ͜͠ͱ͸෩ͷதʹਧ͖ඈ͹ͯ͠͸͍͚ͳ͍

& ୈ࢛ճNGSݱ৔ͷձ اըνʔϜ ͓ՖݟϓϩδΣΫτࢀՃऀͷΈͳ͞· ೔ຊͷσʔλϕʔεΛࢧ͑Δ DBCLS/NBDC/DDBJ ͷΈͳ͞· Bio OSSίϛϡχςΟ BioHackathon/SPARQLthon/PitagoraNetwork
ͷΈͳ͞·

' • ഑ྻղੳͷڭՊॻతͳॻ੶ • Dr. Bonoͷੜ໋Պֶσʔλղੳ (ϝσΟΧϧαΠΤϯε Πϯλʔφγϣφϧ) • NGSͷ࣮ݧख๏΍ղੳͷํ๏ʹ͍ͭͯ஌ΔͨΊͷॻ੶
• ࣍ੈ୅γʔΫΤϯεղੳελϯμʔυʙNGSͷϙςϯ γϟϧΛ׆͔͖͠ΔWET&DRY (༽౔ࣾ) • ౷ܭ΍ػցֶशͷଆ໘͔ΒόΠΦΠϯϑΥϚςΟΫεΛ஌ Γ͍ͨਓ޲͚ • ੜ໋৘ใॲཧʹ͓͚Δػցֶश ଟॏݕఆͱਪఆྔઃܭ (ػցֶशϓϩϑΣογϣφϧγϦʔζ) (ߨஊࣾ) • ࠷৽ͷݚڀൃද΍ߨԋΛYouTubeͰݟΔ • ౷߹TV http://togotv.dbcls.jp/ja/lecture.html • εϥΠυ • speakerdeck.com/inutano

次世代シーケンサーによるメタゲノム解析：桜の花びらに付着した環境DNAを解析する

次世代シーケンサーによるメタゲノム解析：桜の花びらに付着した環境DNAを解析する

More Decks by Tazro Inutano Ohta

Other Decks in Education

Featured

Transcript