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Hurtig In Vivo Fiksering og isolering af translationelle komplekser fra eukaryote celler
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Hurtig In Vivo Fiksering og isolering af translationelle komplekser fra eukaryote celler

Article DOI: 10.3791/62639-v • 14:29 min • December 25th, 2021
December 25th, 2021 •
*1, *1, 2, 2, 1,3, 1
1Division of Genome Sciences and Cancer, The John Curtin School of Medical Research, The Australian National University, 2Centre for Advanced Microscopy, The Australian National University, 3Victor Chang Cardiac Research Institute
* These authors contributed equally

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Vi præsenterer en teknik til hurtigt at stabilisere translationelle (proteinbiosyntese) komplekser med formaldehyd crosslinking i levende gær og pattedyr celler. Tilgangen muliggør dissekering af forbigående mellemprodukter og dynamiske RNA:proteininteraktioner. De krydslinkede komplekser kan bruges i flere downstream-applikationer, såsom i dybe sekventeringsbaserede profileringsmetoder, mikroskopi og massespektrometri.

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Biologi Udgave 178 proteinbiosyntese snapshot capture oversættelse kompleks stabilisering hurtige celleresponser cellulær stress ribosomale komplekser affinitet rensning af oversættelse mellemprodukter translationel kontrol oversættelse kompleks profil sekventering TCP-seq ribosom (oversættelse) profilering ribo-seq ribosom sedimentering ved ultracentrifugering i saccharose gradient
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