Transcript Diapositiva 1 - Docenti.unina.it

Increasing system complexity
Complex biological system: mammalian cell
Cell differentiation program
Circadian rhytms
Cell cycle control
How does cell proliferation works?
Survival
pathway
Proliferation
pathway
Il controllo del ciclo cellulare
Il controllo del differenziamento
Il ritmo circadiano
START
STOP
Ritmo del sonno
Mammals
Cellule
fotorecettive
della retina
con
melanopsina
corticotropina
Surrene
cortisolo
The circadian clock: an example of a complex regulatory system
Pineal gland
18 hpf
AANAT2
Per2 mRNA
What genes are involved?
TRANSCRIPTOME STUDIES
Misure di espressione genica:
1980 Northern Blot/Dot Blot/Differential
tecniche sottrattive /PCR
dysplay e
Northern Blot procedure
For small RNA
Chemical
1-ethyl-3-(3dimethylaminopropyl) carbodiimide
(EDC) cross-linking facilitates the
production of a covalent bond
between the terminal phosphate of
the RNA to a free amino group on
the nylon membrane resulting in
the small RNA becoming tethered
by one end. This allows the full
sequence of the small RNA to be
accessible to hybridizing probes in
contrast to UV cross-linking which
may lead to consumption of uridine
residues during the cross-linking
process and hence compromise the
length of the sequence available
for hybridization .
D box and E BOX binding factors (PAR) and light-induced trancriptional activation
Espressione ubiquitaria o tessuto specifica?
Tissues
Genes
A
B
C
D E F G H I L
Un gene funzionale è evolutivamente conservato: zoo
blot