The central
dogma
Relationships of DNA to mRNA
to polypeptide chain
The basic principle of transcription
The basic principle of translation
Gene mutation and evolution
Gradual accumulation of mutations over long
periods of time results in new biological species,
each with a unique DNA sequence,Mutations
and stabilization of mutations to new species.
Charles Darwin,Unifying evolution theory-the
origin of species from a common ancestor.
Dynamic Cells
Gene mutation and evolution
Gradual accumulation of mutations over long
periods of time results in new biological species,
each with a unique DNA sequence,Mutations
and stabilization of mutations to new species.
Charles Darwin,Unifying evolution theory-the
origin of species from a common ancestor.
A mammalian cell
A plant cell
A bacterial
cell
Endoplasmic reticulum
Golgi apparatus
Mitochondrion
Lysosomes for protein degradation
Nucleus and nucleolus
Chloroplasts in a plant cell
Vacuoles
Biological
motors
-- Dynein
and
kinesin
Cell-surface receptors or channels
Three types of cytoskeletal filaments
Extracellular Matrix
protein
DNA membrane
UV
Normal cell Apoptotic cell
Apoptotic body
stimuli
Programmed Cell Death
Major methods used in biochemistry
? The essence of biochemistry is purification,
reconstitution,and assay readout.
Plasmid vectors for amplification of
DNA inserts
Restriction enzymes
? Viruses can be
used as vectors to
study gene
function
An example of plasmid vectors
Density gradient ultracentrifugation
Size-exclusion chromatography
Ion-exchange chromatography
Transfection
?Preparation of plasmid DNA
?Culture cells to be transfected at a desired
density
?Mix DNA with liposomal transfection reagent,
Liposomal Transfection reagent is used to enclose the
plasmid and fuse with cell membrane,plasmid can then
enter transfect into the cell.
?Add to cell culture; wash the transfection mix
?Let cells grow and express proteins of interest
Immunoprecipitation and kinase assay
Structural biology and bioinformatics
New
drug
design
against
HIV
The scope of proteomics
Protein species
Expression profile
Physical modification
Protein-protein interaction
Enzymatic activity
Two-dimensional gel electrophoresis
? Protein samples separated on an SDS-PAGE gel
Western Blot
? Visualization
of specific
proteins
Western blotting
Principle of western blotting (cont’d)
HRP/hydrogen peroxide catalyzes the oxidation of luminol,
The oxidized luminol is in an excited state,which then decays
to ground state via a light emitting pathway,The emitted light
can be captured by a film.
Principle of western blotting
Protein-protein interactions
Protein-protein interactions
? Protein-protein interactions underlie
virtually all biological events,It has
become a major theme of current
molecular and biochemical studies,
?Yeast two-hybrid screening
?Affinity purification
?Co-immunoprecipitation
Yeast two-hybrid to identify interacting
partners
Range of sizes
of objects
studied by
biochemists
and biologists.
Recent developments in experimental
techniques in biochemistry
Biological Complexity
1,Signal diversity
2,Molecular complexity,thousands of kinases
and phosphatases
3,Pathway diversity
4,Cross-talk of diverse pathways
5,Biodiversity
6,Individuality
Etc….
Cross-talks among research areas
Biochemistry in application
? Disease diagnosis
? Medicine
? Drug design
? Nutrient supplements
? Research reagents
Medical applications of biochemistry
Concluding remarks
? Biochemistry is a biological science on
biomolecules,In other words,the aim of
biochemistry is to understand life in
molecular terms.
? Biochemistry is a multidisciplinary science.
? Again,life is for life.
Thanks
? Thanks to students who continue to teach us
biochemistry by showing us that the best
way to learn a subject is to teach it!
dogma
Relationships of DNA to mRNA
to polypeptide chain
The basic principle of transcription
The basic principle of translation
Gene mutation and evolution
Gradual accumulation of mutations over long
periods of time results in new biological species,
each with a unique DNA sequence,Mutations
and stabilization of mutations to new species.
Charles Darwin,Unifying evolution theory-the
origin of species from a common ancestor.
Dynamic Cells
Gene mutation and evolution
Gradual accumulation of mutations over long
periods of time results in new biological species,
each with a unique DNA sequence,Mutations
and stabilization of mutations to new species.
Charles Darwin,Unifying evolution theory-the
origin of species from a common ancestor.
A mammalian cell
A plant cell
A bacterial
cell
Endoplasmic reticulum
Golgi apparatus
Mitochondrion
Lysosomes for protein degradation
Nucleus and nucleolus
Chloroplasts in a plant cell
Vacuoles
Biological
motors
-- Dynein
and
kinesin
Cell-surface receptors or channels
Three types of cytoskeletal filaments
Extracellular Matrix
protein
DNA membrane
UV
Normal cell Apoptotic cell
Apoptotic body
stimuli
Programmed Cell Death
Major methods used in biochemistry
? The essence of biochemistry is purification,
reconstitution,and assay readout.
Plasmid vectors for amplification of
DNA inserts
Restriction enzymes
? Viruses can be
used as vectors to
study gene
function
An example of plasmid vectors
Density gradient ultracentrifugation
Size-exclusion chromatography
Ion-exchange chromatography
Transfection
?Preparation of plasmid DNA
?Culture cells to be transfected at a desired
density
?Mix DNA with liposomal transfection reagent,
Liposomal Transfection reagent is used to enclose the
plasmid and fuse with cell membrane,plasmid can then
enter transfect into the cell.
?Add to cell culture; wash the transfection mix
?Let cells grow and express proteins of interest
Immunoprecipitation and kinase assay
Structural biology and bioinformatics
New
drug
design
against
HIV
The scope of proteomics
Protein species
Expression profile
Physical modification
Protein-protein interaction
Enzymatic activity
Two-dimensional gel electrophoresis
? Protein samples separated on an SDS-PAGE gel
Western Blot
? Visualization
of specific
proteins
Western blotting
Principle of western blotting (cont’d)
HRP/hydrogen peroxide catalyzes the oxidation of luminol,
The oxidized luminol is in an excited state,which then decays
to ground state via a light emitting pathway,The emitted light
can be captured by a film.
Principle of western blotting
Protein-protein interactions
Protein-protein interactions
? Protein-protein interactions underlie
virtually all biological events,It has
become a major theme of current
molecular and biochemical studies,
?Yeast two-hybrid screening
?Affinity purification
?Co-immunoprecipitation
Yeast two-hybrid to identify interacting
partners
Range of sizes
of objects
studied by
biochemists
and biologists.
Recent developments in experimental
techniques in biochemistry
Biological Complexity
1,Signal diversity
2,Molecular complexity,thousands of kinases
and phosphatases
3,Pathway diversity
4,Cross-talk of diverse pathways
5,Biodiversity
6,Individuality
Etc….
Cross-talks among research areas
Biochemistry in application
? Disease diagnosis
? Medicine
? Drug design
? Nutrient supplements
? Research reagents
Medical applications of biochemistry
Concluding remarks
? Biochemistry is a biological science on
biomolecules,In other words,the aim of
biochemistry is to understand life in
molecular terms.
? Biochemistry is a multidisciplinary science.
? Again,life is for life.
Thanks
? Thanks to students who continue to teach us
biochemistry by showing us that the best
way to learn a subject is to teach it!
