È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
Capitoli
- 00:05Titolo
- 01:21Purification of TcTERT
- 03:18T. castaneum Total RNA Isolation
- 04:53in vitro Reconstitution of T. castaneum Telomerase
- 05:38Telomerase Repeat Amplification Protocol (TRAP) Assay
- 07:57Representative Results for the in vitro Reconstitution of the Active T. castaneum Telomerase
- 08:52Conclusion
Efforts to isolate the catalytic subunit of telomerase, TERT, in sufficient quantities for structural studies, have been met with limited success for more than a decade. Here, we present methods for the isolation of the recombinant Tribolium castaneum TERT (TcTERT) and the reconstitution of the active T. castaneum telomerase ribonucleoprotein (RNP) complex in vitro.
Tags
TelomeraseT. CastaneumCatalytic SubunitIsolationRecombinantTelomerase RibonucleoproteinIn Vitro ReconstitutionDNA RepeatsTelomeresReverse TranscriptaseLinear ChromosomesEnd-to-end FusionDegradationDNA ReplicationHayflick LimitSomatic CellsCancer CellsCell ImmortalityProtein SubunitIntegral RNA ComponentTemplate SynthesisCrystal Structure
