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FACULTAT DE VETERINÀRIA
CENTRE DE BIOTECNOLOGIA ANIMAL
I TERAPIA GÈNICA (CBATEG)
GENETIC MANIPULATION OF THE PANCREAS:
CELL AND GENE THERAPY APPROACHES FOR
TYPE 1 DIABETES
EDUARD AYUSO LÓPEZ

Memòria presentada pel llicenciat EDUARD
AYUSO LÓPEZ per optar al grau de Doctor en
Bioquímica.
Aquesta Tesi Doctoral ha estat realitzada sota la
direcció de la Dra. Fàtima Bosch i Tubert en el
Departament de Bioquímica i Biologia Molecular
de la Facultat de Veterinària i al Centre de
Biotecnologia Animal i Terapia Gènica
(CBATEG).
EDUARD AYUSO LÓPEZ FÀTIMA BOSCH I TUBERT
MAIG de 2006
BELLATERRA

A mis padres, y a mi hermano, por aportar
toda la estabilidad, confianza y ayuda
incondicional imprescindible para realizar este
trabajo.
A Sabrina, por todo lo que hemos superado
juntos, y por todo lo que nos espera en el presente
y el futuro.

Abbreviations
ABBREVIATIONS
AAV Adeno-asociated virus
ABC Avidin-biotin complex acid
Ad Adenovirus
ADP Adenosine-5'-diphosphate
AMPK AMP-activated protein kinase
ATP Adenosine-5'-triphosphate
BAD Pro-apoptotic protein
BM Bone marrow
BMC Bone marrow cells
bp base pair
BSA Bovine Serum Albumin
ß-gal ß-galactosidase
cAMP cyclic adenosine-5'-monophosphate
CAR Coxsackie virus adenovirus receptor
cDNA copied DNA
CAG cytomegalovirus enhancer and chicken beta actin hybrid promoter
Ci curie
CMV cytomegalovirus
Con control
cpm counts per minute
DAB 3’3’-diaminobenzidine
DAG diacylglycerol
dATP desoxiadenosin-5'-triphosphate
DCCT diabetes control and complications trial
dCTP desoxicitidin-5'-triphosphate
DMSO dimethyl sulfoxide
DNA Desoxiribonucleic acid
Dnase deoxyribonuclease
dNTPs 2'-deoxynucleosides 5'-triphosphate
ds double stranded
EDTA ethylenediamine-tetraacetic acid
Fas Fas, cell death receptor
FasL Fas ligand
FBS Fetal Bovine Serum
FG-Ad First generation adenoviral vectors
FITC fuorecein isothiocyanate
g grams
GAD Glutamic acid decarboxilase
GFP Green fluorescein protein
GH Growth hormone

Abbreviations
GK glucokinase
GLP-1 glucagon-like peptide 1
h hour
hCMV human cytomegalovirus
HC-Ad High capacity adenoviral vectors
HD-Ad Helper-dependent adenoviral vectors
HEK Human Embryo Kidney
HIV human immunodeficiency virus
HLA Human leukocyte antigen
HPRT Hypoxanthine-guanine phosphoribosyltransferase
IFN-beta interferon
IFN/IGF-I double transgenic mice RIP/IFN and RIP/IGF-I
IGFBP Insulin-like growth factor binding protein
IGF-I Insulin-like growth factor-I
IGF-II Insulin-like growth factor-II
IGF-IR Insulin-like growth factor receptor-I
IL interleukine
IM Intramuscular
iNOS inducible nitric oxide sinthase
Ip intraperitoneal
IP3 inositol triphosfphate
IR Insulin receptor
IRS-1 Insulin receptor substrate 1
IRS-2 Insulin receptor substrate 2
IU infection units of adenoviral particles
Kb Kilobase
KDa kiloDalton
KO knock-out
M molar (mol/l)
MAPK mitogen-activated protein kinase
MCS Multiple Cloning Site
mg milligrams
MHC major histocompatibility complex
min minute/s
ml millilitre
MLD multiple low dosis
mM millimolar
mRNA RNA messenger
NO Nitric oxide
NOD non obese diabetic
o/n over night
OD optical density
32P Phosphor 32
PBS Phosphate-Buffered Saline
PCR Polymerase Chain Reaction
PDX-1 pancreatic duodenal homeobox gene