Gene Theory
carbon
● Wilhelm Johannsen coined the term
gene in 1909 to name the hereditary
factors referred to by Mendel.
Current definition of gene: nucleic acid
sequence (usually DNA) that encodes for a
functional polypeptide or RNA sequence.
- Genes lead to products essential for the
structure and metabolism of every cell.
One Gene-One Polypeptide
● Beadle and Tatum
- One gene–one polypeptide hypothesis
Gene → Gene product → Phenotypic
expression
- First clear, well-documented study
linking genes and enzymes.
- Define gene more inclusively as a
nucleic acid sequence (usually DNA)
that encodes a functional polypeptide or
RNA sequence.
Storage and Transfer of Genetic Information
Nucleic Acids: Molecular Basis
Inheritance
● Cells contain two kinds of nucleic acids:
deoxyribonucleic acid (DNA) genetic
material and ribonucleic acid (RNA)
which functions in protein synthesis.
DNA and RNA are polymers of nucleotides
- A nucleotide is composed of:
● Pentose sugar with five carbons
(deoxyribose or ribose)
● Nitrogenous base attached to 1′
● Phosphate group attached to 5′
carbon
Two types of nitrogenous bases
Purines
- Contains two fused rings
● Adenine and guanine
● Larger than pyrimidines
● Two fused rings
Pyrimidines
- Characteristic structure is a single,
six-membered ring of carbon and
nitrogen atoms
● RNA: cytosine and uracil
● DNA: cytosine and thymine
● Smaller than purines
● Single hexagonal ring
Backbone of DNA Strands
- The “backbone” of each DNA strand is
alternating phosphate groups and
pentose sugars.
- The 5′ end of each strand has a free
phosphate group attached to the 5′
carbon of the pentose sugar.
- The 3′ end has a free hydroxyl group
attached to the 3′ carbon of the pentose
sugar.
of
Structure of DNA
● DNA is a double stranded molecule
- Strands are joined by hydrogen
bonding:
● Two hydrogen bonds between A
and T
● Three hydrogen bonds between
C and G
- Strands are complementary
● The base sequence in one
strand determines the base
sequence in the other
● Purine base pairs with a
pyrimidine
- Adenine-Thymine
- Guanine-Cytosine
DNA Double Helix
● The DNA strands are antiparallel (two
DNA strands run in opposite directions).
- 5’ end of one is associated with
the 3’ end of the other.
● The DNA ladder is twisted into a double
helix
- Ten base pairs occur per turn
- Each strand is connected by
hydrogen bonds.
DNA Replication
● DNA replication is semiconservative
- DNA molecule separates
- Each parent strand serves as a template
for synthesis of complementary strand.
- Each daughter molecule retains one of
the original parental strands.
● Need several enzymes to accomplish
this:
- Helicase: separates the DNA strands
- DNA polymerase: catalyzes assembly
of the new strand
● Synthesis only occurs in the 5’ to
3’ direction
● Before a cell divides, the structure of
DNA must be precisely copied so that
identical copies can be distributed to the
daughter cells. This is called
replication.
- During replication, the two strands of the
double helix unwind, and each
separated strand serves as a template
against which a complementary strand
is synthesized.
- An enzyme (DNA polymerase)
catalyzes assembly of a new strand of
polynucleotides with a thymine group
going opposite the adenine group in the
template strand, a guanine group
opposite the cytosine group, and the two
reverse conditions.
- DNA polymerase synthesizes new
strands only in the direction of 5' to 3'.
- Because the parent DNA strands are
antiparallel, one of which runs 5' to 3'
and the other running 3' to 5', synthesis
along one of the strands is continuous,
and the other must be formed in a series
of fragments, each of which begins with
a 5' end running toward a 3' end.
Process of DNA Replication
Leading and Lagging Strands
- Since parent DNA strands are
antiparallel.
● One strand is synthesized
continuously
Leading strand
● One strand is synthesized in
fragments, each in the 5’ to 3’
direction
Lagging strand
DNA ligase: joins fragments of the lagging
strand
Transcription of mRNA
● DNA does not directly participate in
protein synthesis.
- The triplet codes of DNA are
transcribed into mRNA; each
triplet (codon) in mRNA encodes
an amino acid.
● Central Dogma of Biology:
DNA->mRNA->protein
- RNA polymerase makes a
complementary copy of one
strand of DNA to mRNA.
- mRNA is a single-stranded
molecule with uracil substituted
for thymine.
- The DNA strand not used as a
template during transcription of
a gene is called antisense
strand.
Modification of mRNA
● mRNA transcribed in the nucleus is
modified before entering the cytoplasm.
Introns removed
- Sections of DNA that do not specify a
part of the finished product.
● Sequence of bases that do not code for
protein.
Exons are spliced together
● Regions encoding part of the mature
mRNA that is translated into protein.
- Before the mRNA leaves the nucleus, a
Methylated guanine cap added to 5’
end, and a Polyadenine tail added to 3’
end.
Translation
● Protein assembly requires several types
of RNA molecules.
- Ribosomal RNA (rRNA)
- Transfer RNA (tRNA)
- Messenger RNA (mRNA)
● mRNA shuttles instructions for protein
synthesis into the cytoplasm from
nucleus.
● Translation takes place on ribosomes.
- Ribosomal RNA (rRNA) contains
a large and a small subunit.
- Subunits fit together to form a
functional ribosome.
 Regulation of Gene Expression
Polyribosomes ● As tissues differentiate, they use only a
● Multiple mRNAs attach to ribosomes part of the genetic instruction present in
forming a complex called a polysome or every cell.
polyribosome. - In a particular cell or tissue
- Each ribosome has one polypeptide ● Genes express
chain. themselves only at
- Allows for multiple copies of a protein to certain times and not at
be synthesized at the same time. others.
● Most genes are inactive
at any given moment.

Gene Regulation in Eukaryotes

● Transcriptional control
- Requires a promoter base
sequence.
- Uses RNA polymerase
- Affected by transcription factors
● Proteins that recognize a
particular promoter
Role of tRNA region, enabling
● The tRNA collects free amino acids and transcription of the
delivers them to mRNA-ribosome attached gene.
complex, which is the site for ● Binding can be affected
polypeptide formation. by variation in base
- A unique tRNA exists for each amino sequences of the
acid. promoter region.
- Each tRNA has a specific tRNA
synthetase (enzyme). Transcriptional Control
● Sorts and attaches an amino ● Also affected by:
acid group to the end of each - Structural genes: encodes for
tRNA called charging. mRNA or rRNA
- A region of the tRNA contains a - Regulatory genes: attached
sequence of 3 bases, the anticodon promoter region affecting where
● Complementary to a codon in structural genes are transcribed.
mRNA. Cis-regulation
- Interaction between structural and
regulatory genes

Trans-regulation
- Genes encoding transcription factors

Methylation of cytosine bases

- Important mechanism for silencing
genes
 • Sticky ends tend to anneal by the rules
of complementary base pairing.
• The ends are later sealed by DNA ligase.

Recombinant DNA
● Recombinant DNA is created when
DNA from a foreign source is ligated
(sealed and joined) into a plasmid.
- Plasmids: small circles of
double-stranded DNA in bacteria (1% to
3%).
● Plasmids are distinct from and
Translational Control much smaller than a bacterial
● Translation is delayed when genes are cell’s primary genome and can
transcribed but mRNA is sequestered. occur in multiple copies per cell.
- Like in oocyte (egg) formation prior to ● To produce recombinant DNA in
fertilization. large quantities. Modified
● Inhibit the expression of, or degrade plasmid must be cloned in
target mRNAs by: treated bacteria cultures.
- miRNAs: short non-coding RNA ● Can carry important genetic
(microRNA) information like resistance to
- siRNAs: small interfering RNA antibiotics.
- Packaged as RISC:
RNA-induced silencing complex Polymerase Chain Reaction (PCR)
● Used to clone known gene sequences
Gene Rearrangement from a variety of organisms with a series
● Rearrangement of DNA sequence of enzymes.
coding for antibodies during lymphocyte - Two short chains of nucleotides
development. called primers are synthesized.
- Results to shuffling of exons encoding ● Primers are
modules of protein function that permits complementary to
the assembly of new and diverse different DNA strands in
antibodies. the known sequences at
- Help combat the wide array of foreign opposite ends of the
antigens in the environment. gene to be cloned.
- At five minutes per cycle, less
Molecular Genetics than two hours is needed to yield
● Restriction endonucleases are a a million copies of one strand.
series of enzymes derived from bacteria.
- Cleave double-stranded DNA at
particular sites determined by base
sequences.
- Cut DNA strands so that one has
several bases projecting farther than the
other strand called-“sticky ends”.
- These DNA fragments are mixed with
others cleaved by the same
endonuclease.