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Tesi etd-10192014-190111


Thesis type
Tesi di laurea magistrale
Author
BISERNI, MARTINA
URN
etd-10192014-190111
Title
Monitoring protein turnover in embryonic stem cells
Struttura
BIOLOGIA
Corso di studi
BIOLOGIA APPLICATA ALLA BIOMEDICINA
Commissione
relatore Prof. Suter, David
relatore Prof.ssa Batistoni, Renata
Parole chiave
  • protein degradation
  • sfGFP
  • protein synthesis
  • fluorescent timer
  • Red fluorescent protein
  • CD-tagging
  • live cell imaging
  • Embryonic stem (ES) cells
  • lentiviral vector
  • proteome
Data inizio appello
11/12/2014;
Consultabilità
completa
Riassunto analitico
Embryonic stem (ES) cells are derived from the inner cell mass of the early embryo (blastocyst) and are able to differentiate into the three primary germ cell layers of the embryo (mesoderm, endoderm and ectoderm) and germ cells. <br>Cells need to change the expression level of the proteins they express in various situations such as during differentiation, cell cycle and in response to environmental cues. Numerous studies have described quantitative changes of the transcriptome, but only few investigated changes at the protein level and do not give information about how the proteome is dynamically regulated. Furthermore, whereas quantitative transcriptome analysis gives indirect hints about protein synthesis, it does not inform about protein degradation. <br>In this project I aimed to monitor dynamics of protein synthesis and degradation in ES cells by tagging endogenous proteins with a fluorescent timer. A fluorescent timer consists of the fusion of two single-color fluorescent proteins with different maturation half times. This allows to monitor not only synthesis but also degradation rate. <br>In the future, this approach will be used to tag several hundreds of different proteins. Interesting patterns of protein synthesis and degradation might be discovered and compared during undifferentiated state versus differentiated state.<br>
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