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Fluorescences in the cytosol: Y-2A-C-2A-P

Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression. CMVp. 2A. EYFP. ECFP. pac. pA. 2A. CMVp. T7p. TRANSLATION. TRANSLATION. 2A. EYFP. ECFP. pac. 2A. wt FMDV 2A. DAPI. pac. ECFP. 2A. EYFP. pA. 2A. T7p. pac. 2A. EYFP.

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Fluorescences in the cytosol: Y-2A-C-2A-P

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  1. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression CMVp 2A EYFP ECFP pac pA 2A CMVp T7p TRANSLATION TRANSLATION 2A EYFP ECFP pac 2A wt FMDV 2A DAPI pac ECFP 2A EYFP pA 2A T7p pac 2A EYFP ECFP 2A mut FMDV 2A DAPI Fluorescences in the cytosol: Y-2A-C-2A-P

  2. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression 2A EYFP GT ECFP pac pA 2A CMVp T7p TRANSLATION 2A EYFP GT ECFP pac 2A DAPI From the cytosol to the Golgi: Y-2A-G-C-2A-P

  3. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression 2A GT EYFP ECFP pac pA 2A CMVp T7p TRANSLATION 2A GT EYFP ECFP pac 2A DAPI ? Problems in the ER translocon: G-Y-2A-C-2A-P

  4. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression G-Y-2A-MT-C-2A-P 2A EYFP MT ECFP pac GT pA 2A CMVp T7p TRANSLATION 2A EYFP MT ECFP pac GT 2A ? Y-2A-MT-C-2A-P 2A EYFP MT ECFP pac pA 2A CMVp T7p TRANSLATION 2A EYFP MT ECFP pac 2A DAPI More problems in the ER translocon: G-Y-2A-MT-C-2A-P

  5. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression G-Y-2A-G-C-2A-P G-C-2A-G-Y-2A-P 2A 2A EYFP GT ECFP pac GT pA ECFP GT EYFP pac GT pA 2A 2A CMVp CMVp T7p T7p TRANSLATION TRANSLATION 2A 2A EYFP GT ECFP pac GT ECFP GT EYFP pac GT 2A 2A ? ? Further problems in the translocon (I): G-Y-2A-G-C-2A-P

  6. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression MT-C G-C-2A-G-Y-2A-P 2A MT ECFP ECFP GT EYFP pac pA GT pA 2A CMVp CMVp T7p TRANSLATION TRANSLATION 2A MT ECFP ECFP GT EYFP pac GT 2A Further problems in the translocon (II): Mitochondria!!

  7. Second course – multicolour mammalian cells: self-processing polyproteins for polycistronic co-expression Type II transmembrane ER-C-2A-G-Y ECFP GT EYFP ER pA 2A CMVp TRANSLATION C ER ECFP GT EYFP ER 2A Cytosol N ? Type I transmembrane secreted ER-C N’ ECFP ER pA 2A CMVp N’ TRANSLATION C ER ECFP ER 2A N N Cytosol C A GalT problem in the polyprotein context?: ER-C-2A-G-Y

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