VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL...
Transcript of VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL...
![Page 1: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/1.jpg)
Erwin R. SchmidtInstitut für Molekulargenetik
3. VL05. 05. 2009
Thema Gentechnologie
![Page 2: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/2.jpg)
Plasmide als Vektoren
![Page 3: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/3.jpg)
Vektor zur Klonierung von PCR-Produkten
![Page 4: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/4.jpg)
Eigenschaften einiger PCR-Polymerasen
Table 1. Comparison of PCR Product Properties for Some Thermostable DNA
Polymerases.
Taq/ VentR®/ DeepCharacteristic AmpliTaq® Tfl Tth (Tli) VentR® Pfu Pwo
>95% >95% Resulting DNA ends 3´ A 3´ A 3´ A Blunt Blunt Blunt
5´®3´ exonucleaseActivity Yes Yes Yes No No No No
3´®5´ exonucleaseactivity No No No Yes Yes Yes Yes
N.A.: not available
![Page 5: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/5.jpg)
Für die Expression fremder Gene gibt es spezielle Expressionsvektoren mit Signalsequenzen für die korrekte Transkription und Translation in Bakterien
![Page 6: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/6.jpg)
Plasmidvektoren, zur Expression von Fremdgenen
![Page 7: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/7.jpg)
![Page 8: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/8.jpg)
![Page 9: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/9.jpg)
![Page 10: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/10.jpg)
Expression von Fusionsproteinen mit sog. „His-tag“
![Page 11: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/11.jpg)
![Page 12: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/12.jpg)
![Page 13: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/13.jpg)
„Gateway“-Klonierungssystem
![Page 14: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/14.jpg)
„Gateway“-Klonierungssystem
![Page 15: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/15.jpg)
„Gateway“-Klonierungssystem
![Page 16: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/16.jpg)
„Gateway“-Klonierungssystem
![Page 17: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/17.jpg)
![Page 18: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/18.jpg)
![Page 19: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/19.jpg)
Replikation M13-Bakteriophage
![Page 20: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/20.jpg)
![Page 21: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/21.jpg)
United States Patent 6803230 Inventors:
Bowdish, Katherine S. (Del Mar, CA) Fredrickson, Shana (Solana Beach, CA)
Wild, Martha (Solana Beach, CA)
![Page 22: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/22.jpg)
![Page 23: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/23.jpg)
![Page 24: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/24.jpg)
![Page 25: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/25.jpg)
Substitutionsvektoren
Linker Arm „Stuffer“ Rechter Arm
IntegratLigation
![Page 26: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/26.jpg)
![Page 27: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/27.jpg)
![Page 28: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/28.jpg)
Sal Bam Eco Eco Bam Sal
Verdau mit Sal I + Eco
Sal Bam Eco Eco Bam Sal
Sal
Sal
Eco
Eco
Sal
Sal
Fällen mit Isopropanol
Eco
Eco
Sal
Sal
Integrat
Linker Arm Rechter Arm„Stuffer“
![Page 29: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/29.jpg)
Lambda Zap:Moderne Vektoren Mit Phagen und Plasmid-eigenschaften
![Page 30: VL Gentechnologie3 09 - molgen.biologie.uni-mainz.demolgen.biologie.uni-mainz.de/Downloads/PDFs/VL Gentechnologie3_09.pdf · Lur scnn 11 en Purified fusion protein Expression Immobilized](https://reader030.fdocuments.us/reader030/viewer/2022041204/5d53f29f88c993a8218b8de3/html5/thumbnails/30.jpg)