Documente Academic
Documente Profesional
Documente Cultură
Introduction to Animal
Biotechnology
Department of Animal Science
National Chung Hsing University
Pin-Chi Tang
Reproductive Biotechnology
Reproductive-biological procedure
AI (artificial insemination)
Estrous synchronization
Regulation of parturition
ET (embryo transfer)
Cryoperservation of gametes and embryos
Sexing of sperm and embryos
IVP (in vitro production of embryos)
Embryo bisection
NT (nuclear transfer)
Microinjection technology (Sperm, DNA, RNA...)4
Estrus Detection
Cow
Swine
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Artificial Insemination, AI
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Advantages of AI
Genetic improvement e.g., widespread use of
outstanding sires; improving accuracy of selection through
progeny test; permitting crossbreeding; introduction of new
genetics
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Cryoperservation of
gametes and embryos
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Cryopreservation
Freezing injury
destroy cell function irreversibly
intracellular ice crystal formation and
salt deposits
changes in the composition of surrounding
milieu at thawing phase
cytotoxicity of cryoprotectants
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Cryoprotective agents
* high water solubility; high permeability;
low toxicity
* alcohols, amines, sugars and proteins
* dimethyl sulfoxide (DMSO); 1,2-propanediol
(PROH); ethylene glycol (EG); glycerol
Antifreeze proteins
* many animals and plants in nature use
cryoprotective agents
* production of antifreeze proteins, mainly
glyopeptides
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Embryo Transfer
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Molecular breeders
Nuclear transfer
Mammalian embryogenesis
by asexual reproduction
Cell transplantation
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Transgenic Animals
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normal mouse
transgenic mouse
Promoter: metallothionine-1
Transgene: rat GH
Science 1982, 385:810-813
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Nuclear transfer
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DNA microinjection
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Enucleation
Nuclear transfer
Nuclear Transfer
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Huntington
s disease (HD)
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Developmental
phase
Production
species
Therapeutic
application
Potential
market
date
AT III
Phase III
Goat
Genetic heparin
resistance
2005
TPA
Phase II / III
Goat
Dissolving
coronary clots
2006
a-AT
Phase II / III
hFVIII
Experimental
sheep
Cystic fibrosis
> 2008
HAS
Phase I
Cattle
Hemophilia A
2008
Various
Abs
Phase I / II
Goat
Blood substitute
> 2007
AT III: antithrombin; TPA: tissue plasminogen; a-AT: antitrypsin; hFVIII; human clotting factor;
Human serum albumin;
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Genomics
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Age of -omics
Genomics: the quantitative study of genes,
regulatory and noncoding sequences.
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Blueprint of Life
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Post-translational
modifications
Functional proteins
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Defining Genes
A complete chromosomal segment
responsible for making a functional product
Open Reading Frame (ORF): a string of codons
bounded by start and stop signals.
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Genome
The entire collection of genes
encoded by a particular organism.
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Mitochondrial genome
Animal has 15-17kb, no intron
Nearly always circular with 37 genes: 13
for proteins, 22 for tRNA and 2 for rRNA.
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Genomics
Genomics is the scientific study of structure,
function and interrelationships of both
individual genes and the genome in its
entirety (Bazer and Spencer, 2005)
The field has evolved from identifying short
nucleotide strings of DNA to the sequencing
of an organism
s complete genome.
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: EST
DNA microarrayproteome
: generation of random mutation
in the genome, ENU, RNAi, T-DNA, EPtransposon, and so on
: generation of target
mutation, knockout
2008 Copyright SYHuang
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http://www.animalgenome.org/
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DNA
(
)(
RN-)
(
)ESRB
MC4R
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ESTs
()
mRNA
(Craig et al., 1990)
DNA
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Proteomics
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mRNA
Protein
Product
Functional
Protein
Product
Translational
Regulation
Transcriptional
Regulation
Post-Translational
Regulation
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Definition of Proteome
The PROTEin complement expressed
by the genOME of an organism
(Wilkins, 1994; Wasinger et al., 1995)
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Definition of Proteomics
The study of protein properties on a
global, integrated view of biological
processes.
Expression Proteomics:
Cell-map Proteomics:
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Samples
Separation
Digestion
Identification
hexokinase
normal samples
diseased samples
enolase
serum, CSF
drug
time-course
ORF YOR213p
MW
IEF
1000s of samples
=3 x1000s of gels
1000s of proteins/gel
106-1012 proteins
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MALDI TOF
MALDI TOF/TOF
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Applications of proteomics
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Drug discovery
* identify disease specific protein
* target validation and signal transduction
* drug mode-of-action
* formal drug toxicology
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* Immunogenetic proteins
# identify immuno-reactivity proteins
# identify allergens
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* Quality control
# bring precision and definition to a new level of
protein-based products
# identify genetic features of a cell line
# detect the presence of a disease in biotechnology
products
2008 Copyright SYHuang
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