Molecular Genetics

Paul Rindler M.S.

 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses
 Discovery of DNA
 1800’s
 Johann Miescher isolated a phosphorous
containing acidic compound from the
nucleus
 1928
 Frederick Griffith demonstrates the
existence of a chemical in bacteria that
caries genetic information
 Discovery of DNA
 1930’s
 Various experiments identify
chromosomes as the source of genetic
information
 Chromosomes are composed of mainly
proteins and deoxyribonucleic acid
(DNA)
 The DNA molecule was considered too
simple to be important so proteins were
thought to carry the genetic information
 Discovery of DNA

 1944
 Oswald Avery showed that degradation
of DNA and not protein resulted in loss
of genetic information
 Discovery of DNA
 Rosalind Franklin
(1951)
 Generated X-ray
crystallography data
suggesting a double
helix with phosphates on
the outside
 Discovery of DNA
 Watson & Crick (1953)
 Used Franklin’s data to develop a model for DNA
 Received a Nobel Prize in 1962
 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses
 DNA Structure

 DNA composition
 Phosphate
 Sugar
 Nucleic acids (bases)
 DNA Structure

 DNA Backbone
 Alternating
phosphates and
sugars
 Bases are
attached to
sugars
 DNA Structure
 DNA is composed of 4 different bases
 Chargraff (1951) data suggested A-T and G-C base
pairing
 Pairs are bonded by hydrogen bonds
 DNA Replication

 Must be completed
prior to cell division

 Helicase enzymes
open the DNA

 Polymerase
enzymes generate
a new DNA strand
on each old
template
 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses
 Gene Expression
 DNA codes for genes
 Genes are expressed as proteins
 Gene expression is regulated
 The genome of every cell in your
body is identical
 Cell type is determined by the genes
the cells express
 Protein Synthesis
 DNA → RNA → Protein
 Transcription
 Translation
 DNA acts as a template for RNA
synthesis
 RNA has uracil (U) rather than thymine
(T)
 Protein synthesis requires 3 types of
RNA
 Messenger (mRNA)
 Protein Synthesis
 Transcription
 mRNA is transcribed from DNA
 Protein Synthesis
 mRNA is read by the synthesis
machinery
 3 nucleotides make up a codon which
codes for 1 of 20 amino acids
 Protein
Synthesis
 Transfer (tRNA)

 Translates mRNA
sequence into
amino acids

 80 nucleotide RNA
molecule

 The anticodon is
specific to 1 of 20
amino acids
 Protein Synthesis
 Ribosomal (rRNA)
 2/3 RNA & 1/3 protein
 2 Subunits
 Coordinates protein synthesis
 Protein Synthesis

 The Genetic Code

 64 possible codons

 1 start codon
(AUG)

 3 stop codons
(UAA, UAG, UGA)
 Protein Synthesis

 Translation
 Initiation
 Elongation
 Terminatio
n
 Protein Synthesis

 Summary
 Transcription
 Translation
 Initiation
 Elongation
 Termination
 Protein Synthesis

 Mutations
 Base
substitution
 Conservative
 Nonconservative

 Insertion/deletio
n
 Frameshift
 Jumping genes
 Mobile elements
 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses
 Biotechnology Toolkit

 Isolation of genes for

study
 Recombinant DNA

 Gene Therapy
 Stem cell research
 Cloning
 Recombinant DNA

 Isolation of genes for study

 Find a gene of interest
 Determine gene sequence
 Express the gene product
 Functional analysis
 Recombinant DNA
 The Tools

 Plasmids
 Small circular DNA molecules
 House sequences of interest

 Bacteria
 Uptake and replicate plasmids
 Express the gene of interest

 Good bang for your buck

 Recombinant DNA

 The Method
 Insert gene of interest into a plasmid
 Bacterial transformation (often E.coli)
 Plasmid replication
 Gene expression
 Protein purification
 Protein analysis
 Recombinant DNA

 Applications
 Determine gene function
 Synthesize useful proteins
 Insulin
 Pest resistance

 DNA mapping
 Gene Therapy
 Gene Therapy
 Stem cell research
 Embryonic stem cells
 Readily available from fertility banks
 Limited government resources

 Ethical concerns

 Compatibility problems

 Adult stem cells

 Uses your own stem cells
 Insufficient technology
 Gene Therapy

 Cloning
 Therapeutic
 Recreate specific body parts
 Too technically challenging

 Reproductive
 Exact replica of an entire organism
 First achieved in 1997 with sheep

 Human success in 2004 but data is

questionable
 Gene
Therapy
 Dolly (1997-2003)
 Scotland 1997
 Nucleus removed
from unfertilized egg
 Nucleus from a sheep
skin cell inserted
 New egg implanted
into surrogate mother
 Died of premature old
age
 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses
 Viruses

 Very simple
 Nonliving organism
 Uses host to
replicate
 2 life cycles
 Lytic
 Lysogenic
 Topics

 Discovery of DNA
 DNA structure and replication
 Gene expression and protein
synthesis
 Biotechnology toolkit
 Viruses