|1910 The term "pleiotropie" is first coined by Ludwig Plate in
He defined pleiotropy as occurring when "several
characteristics are dependent upon ... [inheritance]; these characteristics
will then always appear together and may thus appear correlated".
1930 Ronald Fisher in Geometric Model implies locus
mutations as being capable of affecting essentially all traits.
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
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.
The term 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.
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.
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 (acidic proteins).
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 solubilization.
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
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, one-polypeptide)
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
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.
eukaryotic cells (the kinds of cells found in plants and animals), however,
something very interesting happens before the mRNA leaves the
Some parts of the mRNA are cut away, and the remaining parts
are then stitched back together.
The parts of the mRNA that are cut away
never leave the nucleus, so they are called introns (they stay IN the
Introns regulate the amount of the various proteins that are
"For a 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
The remaining parts that are stitched together are called
exons (they EXit the nucleus).
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 and unwanted 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
His sons, John
Davison Rockefeller (July 8, 1839 May 23, 1937) and
William Avery Rockefeller, Jr. (May 31,
1841 June 24, 1922), were Standard Oil co-founders.
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.
Inducible systems offer researchers the
possibility to deregulate gene expression levels at particular stages of plant
development and in particular tissues of interest.
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 viability.
of a gene can be achieved by a two-step process.
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
Once Cre expression has been switched on and
recombination has occurred, the resultant change in gene expression is, in most
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.
This artificial 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.
This membrane-delocalized translation factor is
inactive unless liberated in the cytosol.
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
However, cisgenesis 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.
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 replication.
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
If 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 responses.
Understanding of the
complexity of the chromatin platform and the epigenetic mechanisms that
contribute to transcriptional control has expanded dramatically in recent
These mechanisms include the presence/absence of histone
modification marks, which form an epigenetic signature to mark active or
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.
In addition, chromatin-remodelling complexes physically alter the
chromatin structure to regulate chromatin accessibility to transcriptional
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
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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