The term "pleiotropie" is first coined by Ludwig
Plate in Festschrift.
He defined pleiotropy as occurring when
"several characteristics are dependent upon ... [inheritance]; these
characteristics will then always appear together and may thus appear
Fisher in Geometric Model implies locus
mutations as being capable of affecting
essentially all traits.
Molecular genetics is
primarily concerned with the inter-relationship between the information
macromolecules DNA (deoxyribonucleic acid)
and RNA (ribonucleic acid) and how these molecules are used to synthesize
polypeptides, the basic component of all proteins.
As the vast majority
of gene expression is dedicated to
synthesis, proteins are the major functional end-points of the DNA template
and account for the majority of the dry weight of a cell.
protein is derived from the Greek proteios, meaning 'of the first rank' and
reflects the important roles of proteins in diverse cellular functions, as
enzymes, receptors, storage proteins, transport proteins, transcription
factors, signaling molecules, hormones, etc.
Proteins are composed of
one or more polypeptide molecules which may be modified by the addition of
various carbohydrate side chains or other chemical groups.
Like DNA and
RNA, polypeptide molecules are polymers consisting of a linear sequence of
repeating units, in this case amino acids.
The latter consist of a
positively charged amino group and a negatively charged carboxylic acid
(carboxyl) group connected by a central carbon
atom to which is attached an identifying
Charged molecules are highly soluble in water.
DNA and RNA are
negatively charged (polyanions) because of the
charges present in their component nucleotides.
Depending on their
amino acid composition, proteins
may carry a net positive charge (basic proteins) or a net negative charge
The hydrogen bonding potential of water molecules
means that molecules with polar groups (including DNA, RNA and proteins) can
form multiple interactions with the water molecules, leading to their
Thus, even electrically neutral proteins are often
readily soluble if they contain an appreciable number of charged or neutral
polar amino acids.
The linear backbone of a
DNA molecule and of
an RNA molecule consists of
alternating sugar residues and organophosphate
Whereas the RNA molecules within a cell normally exist as single
molecules, the structure of DNA is a double helix in which two
DNA molecules (DNA
strands) are held together by weak hydrogen bonds to form a DNA
DNA can adopt different types of helical
Genetic information is encoded by the linear sequence of
bases in the DNA strands (the primary structure).
hydrogen bonding permits RNA-DNA duplexes and double-stranded RNA formation
which are important requirements for gene expression.
Hydrogen bonding can occur between bases
within a single DNA or RNA molecule.
During the process of DNA synthesis
(DNA replication), the two DNA strands of each chromosome are unwound by a
helicase enzyme and each DNA strand directs the synthesis of a complementary
DNA strand to generate two daughter DNA duplexes, each of which is identical to
the parent molecule.
DNA replication is initiated at specific points,
which have been termed origins of replication.
According to their
needs, different cells transcribe different segments of the DNA (transcription
units) which are discrete units, spaced irregularly along the DNA sequence.
Only a portion of the RNA made by transcription is translated into a
The expression of genetic
information in all cells is mostly a one-way system: DNA specifies the
synthesis of RNA and RNA specifies the synthesis of polypeptides, which
subsequently form proteins.
Because of its universality, the DNA >
RNA > polypeptide (protein) flow of genetic information has been described
as the central dogma of gene expression molecular biology.
The shape and
structure of proteins is a crucial aspect of molecular gene expression and
links our understanding of gene expression to the biology of the cell.
While primarily concerned with protein molecules that act on DNA and
RNA sequences, such as transcription factors and histones, the study of gene
expression also focuses on where in the cell expression is modulated.
fact, the modulation of gene expression can occur in the nucleus, the
cytoplasm, or even at the cell membrane due to the impact of proteins on RNA in
those cellular subregions.
In his Nobel lecture, given shortly after he
joined the Rockefeller
Institute for Medical Research, Edward L. Tatum outlined the concepts
fundamental to his one-gene, one-enzyme (understood today as one-gene,
processes in all organisms are under genetic control;
processes are resolvable into a series of individual reactions;
reaction is controlled in a primary fashion by a single gene - 1:1
correspondence of gene and biochemical reaction exists;
mutation of a
single gene results only in an alteration in the ability of the cell to carry
out a single primary chemical reaction.
Geneticist George W. Beadle and
the biochemist Edward L. Tatum were awarded the Nobel Prize largely through the
auspices of the Rockefeller Foundation, for
this hypothesis which turned out to
be entirely false !!!
A cell uses the DNA molecule in the nucleus as
a template for protein
The cell sends a 'messenger RNA' = mRNA into the nucleus
to retrieve the encoding.
The mRNA takes the copied "recipe" out of the
nucleus to the ribosome, which is where proteins are made.
cells (the kinds of cells found in plants and animals), however, something very
interesting happens before the mRNA leaves the nucleus.
Some parts of
the mRNA are cut away, and the remaining parts are then stitched back
The parts of the mRNA that are cut away never leave the
nucleus, so they are called introns (they stay IN the nucleus).
regulate the amount of the various proteins that are being made.
while, geneticists didn't know the purpose of introns, so in typical
evolutionary fashion, many decided that they had no purpose, and introns were
lumped into the category of "junk DNA." As we have learned more about genetics,
we have learned that the evolution-inspired idea of "junk DNA" is, itself,
junk, although some evolutionists still cling to it." - Jay L. Wile
remaining parts that are stitched together are called exons (they EXit the
Each exon represents a "module" of useful
If the cell stitches the exons together in one way, it
makes one protein.
If it stitches the exons together in another way, it
makes a different protein.
As a result, a single gene can actually produce
many different proteins.
molecular biology is based on this
Genetically modified organisms
and the industrial manipulation of molecular genetics is based on a theory that
has turned out to be entirely false.
Unintended toxic proteins is the
transgenic standard !
We should not be surprised at
Rockefeller's hand in this as John D. Rockerfeller's father made his fortune
selling "patent" medicine.
William Avery "Bill" Rockefeller, Sr.
(November 13, 1810 May 11, 1906) was
an American confidence
man who went by the alias of Dr. William Levingston.
He worked as a
traveling salesman who identified himself as a "botanic physician" and sold
John Davison Rockefeller (July 8,
1839 May 23, 1937) and William Avery
Rockefeller, Jr. (May 31, 1841 June 24, 1922), were
Several systems for
induction of transgene expression in
plants have been described recently.
Inducible systems were used mainly
in tobacco, rice,
Arabidopsis, tomato, and corn.
systems offer researchers the possibility to deregulate gene expression levels
at particular stages of plant development and in particular tissues of
The more precise temporal and spatial control, obtained by
providing the transgenic plant with the appropriate chemical compound or
treatment, permits analysis of the function of those genes required for plant
Specific mutation of a gene can be achieved by a two-step
Introduction of loxP sites around a functionally essential
genomic part followed by a cell type-specific Cre recombinase-mediated excision
of the loxP flanked sequence.
The same strategy can be used for cell
type-specific overexpression of a transgene, when a strong overall expressing
promoter is separated from the coding region of a gene of interest by loxP
flanked 'STOP' sequences.
In both scenarios, a Cre recombinase
transgene provides spatial control.
Once Cre expression
has been switched on and recombination has occurred, the resultant change in
gene expression is, in most cases, irreversible.
In the quest for the
development of pharmacological switches that control gene expression, no system
has been reported that regulates at the translational level but several systems
have been constructed:
-Tetracycline-inducible transgenic systems
[tetracycline transactivator (tTA) or 'Tet-Off' and reverse tetracycline
transactivator (rtTA) or 'Tet-On'] allow for reversible temporal regulation of
transgene expression .
Between these, rtTA is better suited for rapid
induction of gene expression.
-To permit small-molecule control of
transgene translation a farnesyl transferase inhibitor-responsive translation
initiation factor was constructed.
protein is a three-component chimaera consisting of the ribosome
recruitment core of the eIF4G1 eukaryotic translation initiation factor, the
RNA-binding domain of the R17 bacteriophage coat protein and the plasma
membrane localization CAAX motif of farnesylated H-Ras.
membrane-delocalized translation factor is inactive unless liberated in the
Farnesyl transferase inhibitor FTI-277 prevents the membrane
association of the CAAX motif and thus increases the cytoplasmic levels of the
eIF4G fusion protein, which is then capable of inducing translation of the
second cistron of a bicistronic messenger RNA containing an R17-binding site in
its intercistronic space.
- The concept of cisgenesis could become a
promising approach in future apple breeding.
depends on the availability of effective tools for the
production of marker-free
genetically modified (GM) plants.
The development of such plants was
recently shown to be possible using a heat shock inducible Flp/FRT recombinase
based transformation system allowing the excision of the marker gene from the
genome of GM apple plant tissue.
- A new laser mediated
method heat shocks
groups of cells allowing precise spatio-temporal control of gene expression
without requiring knowledge of specific enhancer sequences.
baculovirus Autographa californica nucleopolyhedrovirus (AcNPV) has been widely
used to achieve a high level of foreign gene expression in insect cells, as
well as for efficient gene transduction into mammalian cells without any
In addition to permitting efficient gene
delivery, baculovirus has been shown to induce host innate immune responses in
various mammalian cells and in mice.
The major barrier to the clinical application of adenovirus gene
therapy for diseases that require stable transgene expression is the
immunogenicity of recombinant adenovirus, which ordinarily limits the duration
of its effects to a period of about 2 weeks.
tolerance to adenovirus
could be induced then transgene expression could be prolonged if
T lymphocytes underwent
thymic selection in the presence of adenovirus antigens.
The ability to
achieve unresponsiveness to a recombinant adenovirus after inoculation of the
thymus in neonates extends the
paradigm of intrathymic tolerance induction.
T lymphocytes are exquisitely
poised to respond rapidly to pathogens and have proved
an instructive model for
exploring the regulation of inducible genes.
Individual genes respond
to antigenic stimulation in different ways, and it has become clear that the
interplay between transcription factors and the chromatin platform of
individual genes governs these
Understanding of the complexity of the chromatin
platform and the epigenetic
mechanisms that contribute to transcriptional control has expanded
dramatically in recent years.
These mechanisms include the
presence/absence of histone modification marks, which form an epigenetic
signature to mark active or inactive genes.
These signatures are
dynamically added or removed by epigenetic enzymes, comprising an array of
histone-modifying enzymes, including the more recently recognized
chromatin-associated signalling kinases.
chromatin-remodelling complexes physically alter the chromatin structure to
regulate chromatin accessibility to transcriptional regulatory factors.
The advent of genome-wide technologies has enabled characterization of
the chromatin landscape of T lymphocytes
in terms of histone occupancy, histone modification patterns and transcription
factor association with specific genomic regulatory regions, generating an
image of the T lymphocyte epigenome.
The use of transgenic
animals in the field of molecular genetics is standard and neccesary for
Molecular genetics experiments use only model
organisms with known regulatory DNA sequences, i.e. enhancers, that drive gene
expression at particular times in development and in particular cells.
By reducing the variables
it becomes easier to predict mutational effects with transgenic animals.
It is impossible to
predict how these genetic switches would function in animals outside of those
specifically bred for the purposes of these experiments.
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