GeneArt - PDF Free Download

GeneArt

... 2 GeneArt...... 3 GeneArt... 4 GeneArt Gene Synthesis... 5 GeneArt Strings DNA Fragments Libraries... 7... 9... 11 &... 12 GeneArt Elements Vector Construction... 13 GeneArt Elements Combinatorial Parts Assembly... 15 Gene-to-protein... 17 GeneArt Directed Evolution... 18 GeneArt... 19

GeneArt PCR PCR Synthetic biology GeneArt GeneArt 1 Invitrogen Vector NTI Ion Torrent SOLiD 2 Invitrogen GeneArt GeneOptimizer 3 GeneArt 4 www.thermofisher.com/genesynthesisproductselection 2

GeneArt... GeneArt Gene Synthesis 100 DNA Invitrogen GeneArt Gene Synthesis ISO 9001 2008 GeneArt Strings DNA Fragments PCR 3kbp DNA Invitrogen GeneArt Strings DNA Fragments GeneArt GeneArt GeneArt GeneArt Directed Evolution Directed Evolution Invitrogen GeneArt Directed Evolution ISO 9001 2008 ISO 13485 2003 3

GeneArt GeneArt Global Centers of Excellence 1999 GeneArt 1 6.5Mbp 1 1.5Mbp ISO 300 4

GeneArt Gene Synthesis GeneArt Gene Synthesis DNA 100 DNA 6 1 GeneArt Gene Synthesis mrna Ready-to-use ISO 100 Web GeneOptimizer 1,200bp 9 * 3,000bp 12 1,200bp 7 3,000bp 10 SuperSPEED** 1,200bp 5 1,800bp 7 * 5 1 ** SuperSPEED TEL 03-6832-6980 1 2 jpgene@liftech.com 5

GeneArt Gene Synthesis GeneOptimize 最適化アルゴリズム発現 GeneAssembler 人工遺伝子合成プラットフォーム野生型 GeneArt DNA 配列 GeneOptimizer により最適化された配列 Classic cloning A C G C G? cdna ライブラリ PCR 増幅クローニング部位の付加ライゲーション & 形質転換コロニーのスクリーニングシーケンシングプラスミドの完成 1. GeneArt Gene Synthesis 1 8Mbp DNA 10kb kb DNA 25 DNA QC www.thermofisher.com/longgenes 6

GeneArt Strings DNA Fragments Libraries Invitrogen GeneArt Strings DNA Fragments 150bp~ 3,000bp のーンの DNA ー GeneArt Gene Synthesis オオ ( 2) Strings DNA Fragments のーンのーンのののーン Strings DNA Fragments のーンのラの ( 3 のーン ) DNA ライラーの Invitrogen GeneArt Strings DNA Libraries ーー Strings DNA Fragments ー GeneArt Strings DNA Libraries オ 3 200~2,000 bp の GeneArt StringsDNA Fragments のー 30 bp IUPAC ー 200ng のの Strings DNA Fragments 9 ( ~ 1,000bp) 12 ( 1,001~3,000bp) * * の加 GeneArt ーイオンラインのインのーン ( ーン TOPO ーン Gateway ーン GeneArt Seamless Cloning and Assembly GeneArt Type IIs Assembly のーン ) Strings DNA Fragments オ DNA PCR DNA の DNA ランーのダイーン 90% ののーンのイライン GeneArt Strings DNA fragments ~1 kb GeneArt Strings DNA fragments 1~2 kb GeneArt Strings DNA fragments 2~3 kb 2 ~ 4 のーンーン 3 ~ 5 のーンーン 4 ~ 8 のーンーン Ordering information 製品名 GeneArt Strings DNA Fragments ( オンラインオーダー, >200 ng ) Email オーダー時の追加料金 * 1 納期 * 2 150 ~ 600 bp 約 9 営業日 ~ 601 ~ 750 bp 751 ~ 1,000 bp 1,001 ~ 1,250 bp 1,251 ~ 1,500 bp 1,501 ~ 1,750 bp 価格 * 3 納期 * 2 約 12 営業日 ~ 1,751 ~ 2,001 ~ 2,000 bp 2,250 bp 2,251 ~ 2,500 bp 2,501 ~ 2,750 bp 2,751 ~ 3,000 bp 13,200 16,500 19,800 42,900 49,500 55,000 60,500 66,000 71,500 77,000 82,500 1,200 1,500 1,800 3,900 4,500 5,000 5,500 6,000 6,500 7,000 7,500 *1 E-mai(l Excel イ ) 時追加料金 *2 時時の時 *3 の 2016 4 のの www.thermofisher.com/strings 7

遺伝子設計 GeneArt テクノロジーを用いた DNA 断片の作製 G A G A 5 5 5 5 5 5 5 5 5 5 5 5 5 5 A G G オリゴヌクレオチドの合成遺伝子アセンブリ直接クローニング直接アセンブリ下流アセンブリ 2. GeneArt Strings DNA Fragments 120% Cloning efficiency Fidelity 100% 100 100 100 94 80% 88 88 86 80 75 88 88 75 83 60% 50 50 67 50 63 63 56 40% 20% 0 624 698 844 960 984 2,545 2,725 2,894 2,946 2,980 断片サイズ ( bp) 3. GeneArt Seamless Cloning and Assembly Kit 3,000bp 10 Strings DNA Fragments PCR fidelity 8

GeneOptimizer 4 100 Fath et al. 2011 GeneOptimizer mrna mrna GC mrna RNAi RNA-resitant form 9

4. GeneOptimizer 100 GeneOptimizer mrna 5 Fath Fath et al. 2011 Mock Wild type GeneArt GeneOptimizer Competitor 1 Competitor 2 Competitor 3 Competitor 4 Competitor 5 25 kda 15 kda 70 kda 55 kda Protein 1 Standardized protein load 5. -His 2 3 Stephan Fath, Asli Petra Bauer, Michael Liss, Anne Spriestersbach, Barbara Maertens, Peter Hahn, Christine Ludwig, Frank Schafer, Marcus Graf, and Ralf Wagner, Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression, PLOS One 6: 3, e17596. www.thermofisher.com/geneoptimization 10

expression-ready 4-5 6 GeneArt DNA pmx pmx & 11 & Express Service 2 Super SPEED 7 4kbp GeneArt GeneOptimizer pcdna 3.1(+) pfastbac 1 pet151/d-topo pcdna 3.3-TOPO pcdna 3.4-TOPO pdonr 221 pet100/d-topo prset A pyes2.1/v5-his-topo www.thermofisher.com/expressgenes GeneArt Gene Synthesis オンラインポータル GeneOptimizer software GeneAssembler process 人工遺伝子合成 :9 営業日 pcdna 3.1* エクスプレスクローニング :1~2 営業日 GeneArt Gene Synthesis GeneOptimizer software GeneAssembler process pmx pcdna 3.1* オンラインポータル人工遺伝子合成 + pmx ベクターへのクローニング :9 営業日サブクローニング : 5~6 営業日 6. pmx * www.thermofisher.com/expressgenes 11

& GeneArt GeneArt Invitrogen GeneArt Elements GeneArt ready-to-use 100 DNA DNA 0.01EU µg pdna pdna CoA DIN ISO GeneArt GeneArt DNA DNA www.thermofisher.com/geneartplasmid 12

GeneArt Elements Vector Construction Invitrogen GeneArt Elements DNA Elements GeneArt in vitro 7 7. GeneArt GeneArt Elements Vector Construction 13

GeneArt Elements DNA * 100 GeneArt CAD Ig tpa GFP Fc-Tag HEK 293 GFP 8 GeneArt Elements Mock pcmv-igk-leader+emgfp pcmv-tpa-leader+emgfp pcmv-gfp (cellular) GFP-Fc GFP 8. GeneArt Elements pcmv-gfp PCML-GFP GFP * GeneArt Elements www.thermofisher.com/elementsvc 14

GeneArt Elements Combinatorial Parts Assembly Combinatorial Parts Assembly CPA DNA 9 DNA CPA ISO 9001 2008 9. CPA 5 5 15

CPA Invitrogen GeneArt Elements 10 TEF1 TEF1 2 桁の違い 10. 9 CPA 25 www.thermofisher.com/elementscpa 16

Gene-to-protein Gene-to-proteins GeneArt DNA CHO FreeStyle 293 FreeStyle CHO Expi293 Sf9 Sf9 FreeStyle FreeStyle CHO Expi293 293 SDS-PAGE 1L L His GST Fc www.thermofisher.com/g2pservice 17

GeneArt Directed Evolution GeneArt Directed Evolution DNA TRIM 50 60 70 80 wild type peer A01 peer A02 peer A03 peer A04 GVVPILVELDGDVNGHKXXVSGXGEXXATYGKLTLKFICT GVVPILVELDGDVNGHKRQVSGGGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKAGVSGEGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKIYVSGLGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKLKVSGPGEGDATYGKLTLKFICT mrna PCR 2µg DNA 10 9 30µg DNA 12x 0.5mL 1 DNA 5 40bp 100 5µg 50 60 70 80 wild type F58A F59A F60A F61A GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKASVSGEGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKFAVSGEGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKFSASGEGEGDATYGKLTLKFICT GVVPILVELDGDVNGHKFSVAGEGEGDATYGKLTLKFICT www.thermofisher.com/directedevolution 18

GeneArt GeneArt Gene Synthesis "GeneArt Portal" GeneArt in silico GeneArt Project Manager 11 Project Configurator 12 11. GeneArt Project Manager 12. GeneArt Project Configurator www.thermofisher.com/genesynthesis 19

GeneArt 13 GeneArt GeneArt Invitrogen GeneArt Strings DNA Fragments Invitrogen GeneArt Elements GeneArt Precision TALs GeneArt Directed Evolution GeneObserver 24 & pmx 5 3 13. GeneArt GeneArt www.thermofisher.com/genearttutorials 20

Notes

www.thermofisher.com/geneart www.thermofisher.com/tc For Research Use only.not for use in diagnostic procedures. 2016 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Printed in Japan. IVC048-A1603OB 108-0023 4-2-8 0120-477-392 jptech@thermofisher.com TEL 03-6832-6980 FAX 03-6832-9584 TEL 03-6832-9300 FAX 03-6832-9580 facebook.com/thermofisherjapan @ThermoFisherJP www.thermofisher.com