Basic Mendelian Genetics

Dr. E. Hannig
BIOL3301_Fall 2013

History of Genetics
4000 years ago, Egyptians practiced animal and plant breeding.essentially
recognizing that desirable traits could be inherited and bred into various plants and
animals.
Year

Scientist(s)
Charles Darwin
1858 Alfred Russel
Wallace
1859 Charles Darwin
1866 Gregor Mendel
Carl Correns
Hugo de Vries
1900
Erich von
Tschermak
1902
1905

1905

1908

1910

1927
1928

1931

Discovery
Joint announcement of the theory of natural selection-that
members of a population who are better adapted to the
environment survive and pass on their traits.
Published The Origin of Species.
Published the results of his investigations of the inheritance of
"factors" in pea plants.
Mendel's principles were independently discovered and
verified, marking the beginning of modern genetics.

Pointed out the interrelationships between cytology and

Walter Sutton
Mendelism, closing the gap between cell morphology and
heredity.
Nettie Stevens
Independently described the behavior of sex chromosomesEdmund Wilson XX determines female; XY determines male.
W Bateson gives the name genetics (means 'to generate' in
Greek) to this branch of science, and introduces the words
allele (allelomorph), heterozygous (impure line) and
Wm. Bateson
homozygous (pure line); W Bateson & RC Punnett work out
the principles of multigenic interaction (linkage) and heredity
Proposed that some human diseases are due to "inborn errors of
Archibald Garrod
metabolism" that result from the lack of a specific enzyme.
Proposed a theory of sex-linked inheritance for the first
Thomas Hunt
mutation discovered in the fruit fly, Drosophila, white eye
Morgan
[Genes reside on Chromosomes]. This was followed by the
gene theory, including the principle of linkage.
Hermann J.
Used x-rays to cause artificial gene mutations in Drosophila.
Muller
Proposed that some unknown "principle" had transformed the
Fred Griffith
harmless R strain of Streptococcus pneumoniae to the virulent
S strain.
Harriet B.
Creighton
Demonstrated the cytological proof for crossing-over in maize.
Barbara
[Crossing over is responsible for recombination]
McClintock

George Beadle
Edward Tatum
Oswald Avery
1944 Colin MacLeod
Maclyn McCarty
1941

Irradiated the red bread mold, Neurospora, and proved that the
gene produces its effect by regulating particular enzymes.
Reported that they had purified the transforming principle in
Griffith's experiment and that it was DNA.

Organized a phage course at Cold Spring Harbor Laboratory

which was taught for 26 consecutive years. This course was the
1945 Max Delbruck
training ground of the first two generations of molecular
biologists
late Barbara
Developed the hypothesis of transposable elements to explain
1940s McClintock
color variations in corn.
Discovered a one-to-one ratio of adenine to thymine and
1950 Erwin Chargaff guanine to cytosine in DNA samples from a variety of
organisms.
1951 Rosalind Franklin Obtained sharp X-ray diffraction photographs of DNA.
Used phages in which the protein was labeled with 35S and the
Martha Chase
DNA with 32P for the final proof that DNA is the molecule of
1952
Alfred Hershey
heredity.
Francis Crick
Solved the three-dimensional structure of the DNA molecule.
1953
James Watson
Matthew
Used isotopes of nitrogen to prove the semiconservative
1958 Meselson
replication of DNA.
Frank Stahl
Purified DNA polymerase I from E. coli, the first enzyme that
1958 Arthur Kornberg
made DNA in a test tube.
Marshall
Nirenberg
Led teams that cracked the genetic code- that triplet mRNA
1966
H. Gobind
codons specify each of the twenty amino acids.
Khorana
Hamilton Smith Isolated the first restriction enzyme, HindII, that could cut
1970
Kent Wilcox
DNA molecules within specific recognition sites.
Paul Berg
1972
Produced the first recombinant DNA molecules.
Herb Boyer
Led the team at Cold Spring Harbor Laboratory that refined
1973 Joseph Sambrook DNA electrophoresis by using agarose gel and staining with
ethidium bromide.
Annie Chang
Showed that a recombinant DNA molecule can be maintained
1973
Stanley Cohen
and replicated in E. coli.
International meeting at Asilomar, California urged the
1975
adoption of guidelines regulating recombinant DNA
experimentation.
Developed the chain termination (dideoxy) method for
1977 Fred Sanger
sequencing DNA.
2

Maxam and Gilbert report chemical DNA sequencing method.

The first genetic engineering company (Genentech) is founded,
using recombinant DNA methods to make medically important
drugs.
Somatostatin became the first human hormone produced
using recombinant DNA technology.

1977
1978

JW Gordon et al produce the first transgenic mouse; Dr Chakrabarty

is awarded the first patent for a genetically engineered (unicellular)
organism; GD Snell, J Dausset, and B Benacerraf receive the Nobel
prize for their studies on the MHC locus

1980
1981
1983 James Gusella
1985 Kary B. Mullis
1988

1989 Alec Jeffreys

1989

1990

1993

Francis Collins
Lap-Chee Tsui

Three independent research teams announced the discovery of

human oncogenes (cancer genes).
Used blood samples collected by Nancy Wexler and her coworkers to demonstrate that the Huntington's disease gene is on
chromosome 4.
Published a paper describing the polymerase chain reaction
(PCR), the most sensitive assay for DNA yet devised.
The Human Genome Project began with the goal of
determining the entire sequence of DNA composing human
chromosomes.
Coined the term DNA fingerprinting and was the first to use
DNA polymorphisms in paternity, immigration, and murder
cases.
Identified the gene coding for the cystic fibrosis
transmembrane conductance regulator protein (CFTR) on
chromosome 7 that, when mutant, causes cystic fibrosis.
First gene replacement therapy-T cells of a four-year old girl
were exposed outside of her body to retroviruses containing an
RNA copy of a normal ADA gene. This allowed her immune
system to begin functioning.
FlavrSavr tomatoes, genetically engineered for longer shelf
life, were marketed.

1997

Complete Saccharomycetes cerevisiae genome is sequenced; complete

E.coli genome is sequenced [Science 277;1453-74]

1998

Caenorhabditis elegans becomes the first animal whose genome is

totally sequenced [Science 282:2012-8]

2000

The Human Genome Project presents its preliminary results: each

of the body's 100 trillion cells contains some 3.1 billion nucleotide
units. Only 1% of these are thought to be transcriptional, clustered
in possibly as few as 30,000 genes.

2003

Successful completion of the Human Genome Project [99%

sequenced with 99.9% accuracy]

2005

Next generation DNA sequencing; massively parallel sequencingby-synthesis; multiplex sequencing; four color sequencing-bysynthesis.

2007 Fred Sanger

Large scale DNA sequencing

2008

Single molecule DNA-sequencing

2010

Single molecule real time DNA sequencing

[modified from http://www.biography.com/people/gregormendel-39282

Gregor Mendel [1882-1884]

1843- Mendel began studying to be a monk: He joined the
Augustinian order at the St. Thomas Monastery in Brno,
and was given the name Gregor.
1849- sent to fill a temporary teaching position in Znaim,
but failed a teaching-certification exam the following year,
1851-sent to the University of Vienna, at the monasterys
expense, to continue his studies in the sciences. Mendel
studied mathematics and physics under Christian Doppler,
after whom the Doppler effect of wave frequency is named; he studied botany under
Franz Unger, who had begun using a microscope in his studies, and who was a proponent
of a pre-Darwinian version of evolutionary theory.
1853-completed his studies at the University of Vienna, returned to the monastery in
Brno and was given a teaching position at a secondary school, where he would stay for
more than a decade. It was during this time that he began the experiments for which he is
best known [these continued until ~1863]
1854-Mendel began to research the transmission of hereditary traits in plant hybrids. At
the time of Mendels studies, it was a generally accepted fact that the hereditary traits of
the offspring of any species were merely the diluted blending of whatever traits were
present in the parents. It was also commonly accepted that, over generations, a hybrid
would revert to its original form, the implication of which suggested that a hybrid could
not create new forms.

1865-Mendel delivered two lectures on his findings to the Natural Science Society
in Brno, who published the results of his studies in their journal the following year, under
the title Experiments on Plant Hybrids. Mendel did little to promote his work, however,
and the few references to his work from that time period indicated that much of it had
been misunderstood. It was generally thought that Mendel had shown only what was
already commonly known at the timethat hybrids eventually revert to their original
form. The importance of variability and its evolutionary implications were largely
overlooked. Furthermore, Mendel's findings were not viewed as being generally
applicable, even by Mendel himself, who surmised that they only applied to certain
species or types of traits.

Other theories of inheritance

Pangenesis
o e.g., Socrates, Charles Darwin
o body parts produce gemmules that are collected in the semen
o similar to earlier arguments by Aristotle and Hippocrates (hereditary
material collects from throughout the body, transmitted via semen and
menstrual blood, interact in the womb to influence early development
Blending theory of Inheritance
o and gametes contain essences that are combined in the offspring
o for example, red flower x white flower gives pink offspring

..vs.

Particulate theory of Inheritance

o genetic elements passed to offspring as discrete elements
o Mendels approach was critical, as he applied a quantitative elements
to his studies
o (i.e.,as do modern geneticists, he counted things!)

Mendels Model System: The Garden Pea (Pisum sativvum)

Why Pisum sativvum?

o access to varieties with
different traits
o self pollination and
hybrid formation
possible
Stamen is the maleorgan,
consists of anther at the tip of
the filament
Pistel consists of a sticky stigma at the tip, connected to the ovary via the
style
Keel encloses both anthers and ovaries
Fertilization:
o Stamens drop pollen in the immature flower, falls on the stigma.
o Pollen tube grows down the style (stalk) to the ovary
o Ovary contains ovules, each of which contains a single egg cell
o Eggs are fertilized, ovule develops into seed.

Diversion..The garden pea was Mendels model system. Other model systems for
genetic studies include:

Saccharomyces
cerevisiae
(budding/bakers yeast)

Drosophila melanogaster
(fruit fly)

Caenorhabditis elegans (nematode)

Zea mays

Danio rerio (zebrafish)

What are the characteristics of a good model system?

o access to varieties with different traits
o short generation times
o inexpensive to grow/keep
o take up little space
o ability to make controlled crosses
o each generation gives rise to a large number of offspring

Mendels Seven Traits (Phenotypes)

Establishment of True-breeding lines:
-Mendel chose traits that showed discontinuous variation (i.e., relatively
discrete polymorphisms) Why??
Mendel selfed (i.e., selfpollinated) plants for 1-2 years
in order to establish lines that
were true-breeding or each
character. [What does this
mean??]

-These are termed pure lines

and, aside from counting, is the
single most important thing
Mendel did that helped
establish his theories!

The Monohybrid Cross

Experiment:
cross [i.e., form a hybrid] a
true-breeding purpleflowered plant with true
breeding white flowered
plant [these represent the P
or parental generation]
Result:
All of the F1 plants (first filial
generation) expressed the
purple flower phenotype.
Identical results were
obtained with a reciprocal
cross [What is a reciprocal
cross??]
Note that in order to determine the F1 phenotype, the fertilized plant must be
allowed to mature to seed pods, the seeds removed, and planted to observe
the flower color of the F1 progeny [Seed color and shape are observed
directly in the mature seed pod and do not require planting the seeds to
observe the F1 phenotype]

For each of the traits, monohybrid crosses of true-breeding strains with

opposing phenotypes yielded F1 expressing only one of the two possible
phenotypes.

-In other words, one of the two phenotypes disappeared in the F1 offspring!
-The trait that remained (i.e., the trait expressed in the F1) came to be know
as the Dominant trait
-The trait that disappeared in the F1 generations came to be known as the
Recessive trait.

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Next Step: Selfing the F1 Generation

3:1 ratio [dominant trait : recessive trait]

observed in the F2 generation from
selfing the F1 plants

Trait

Mendels F1 selfing cross

Cross
F1

F2

F2 ratio

In each case, selfing the F1 plants (which expressed only the dominant
trait) yielded an F2 generation in which the recessive trait
reappeared.but in a specific 3:1 ratio in all cases!
-therefore, the particular determinant for the recessive trait did not disappear
(or no longer exist) in the F1it just wasnt expressed, perhaps masked
by the presence of the dominant particulate element.

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Upon selfing the F2 generation, Mendel discovered that:

1. All of the F2 progeny expressing the recessive phenotype were truebreeding.**
2. F2 progeny expressing the dominant phenotype were not all equal
-1/3 of the dominant F2 plants were true-breeding for the
dominant phenotype
-2/3 of the dominant F2 plants, when selfed, yieled F3 progeny
in the same 3:1 ratio as seen in the F2 generation [in other
words, these 2/3 expressing the dominant trait appeared
genetically identical to the F1 plants!!]

[How to read the figure above: the F2 column represents the 3:1 ratio of
plants seen in the F2 generation. Following the arrows, each row represents
the F3 obtained by selfing the each of the F2 plants]
Underlying the 3:1 phenotypic ratio in the F2 was a 1:2:1 genotypic ratio

**Refer back to when Mendel was initially establishing lines of plants that were truebreeding for each of the traits he studied.

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Based on these observations [remember, DNA, RNA, the nature of genes

were unknown at that time.chromosomes were know, but the relationship
to Mendels elements had not yet been established!!], Mendel deduced that:
1. There existed 2 forms of the hereditary determinant for each trait [1
gene (e.g., flower color), 2 forms..alleles.of each gene (purple,
white)]. Each parent harbored two determinants that may either be
identical or not.
2. Each gamete contains only one of the two determinants in each parent,
such that the offspring has 2 determinants (1 from each parent, united
in the zygote) [What was the alternative explanation??]
3. The probability of either of the 2 determinants in each parent
appearing in a gamete is equal. This is Mendels First Law: The
Principle of Segregation.

The union of male and female gametes in the zygote (offspring) is also a
random process.

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Putting it all together

14

..and a more specific example

15

Selfing the F2 generation unmasks the 1:2:1 genotypic ratio present in

the 3:1 phenotypic ratio.
o of the of the F2 expressing the dominant phenotype, 1/3 were truebreeding for the dominant trait
o of the of the F2 expressing the dominant phenotype, 2/3 were NOT
true-breeding; when selfed, these 2/3 yielded the same 3:1 ratio seen
in the F2 generation [suggesting their genetic makeup was identical
with that of the F2 generation].
o of the of the F2 expressing the recessive trait, ALL were truebreeding for the recessive trait.

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Without knowing about DNA, RNA, Molecular Genetics, etc, Mendel was
able to deduce [based on his studies]:
1. There are 2 hereditary determinants for each trait studied [i.e., each
parent two copies of the determinant for flower color, etc.] [in modern
terms, 1 gene, 2 different alleles]
2. Each gamete contains only one of the two determinants present in
each parent. The probability of either appearing in a gamete is equal
[Mendels First Law, the principle of segregation]
3. The union of male and female gametes in the zygote reunites the
hereditary determinants in pairs. This is also a random process.

Based on these data, Mendel also deduced that in true-breeding

strains, both alleles were identical (homozygous).

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Mendels Second Law: Principle of Independent Assortment

During gamete formation, the segregation of one gene pair is independent of
the segregation of other gene pairs**
The multiplicative rules of probability apply to a cross involving 2 or more
gene pairs that assort independently from each other

What is the ratio of RY : Ry : rY : ry? [Assume R and Y are completely

dominant over r and y, respectively]
Note that, in the above data, R:r and Y:y each segregate in a 3:1 ratio!

**he was lucky, of course, as this does not apply to linked genes, which do
not assort independently.

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Studied by means of the dihybrid cross

The 9:3:3:1 ratio above is characteristic of a dihybrid cross involving 2

genes that assort independently

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For a cross of AaBb x AaBb

o If A and B assort independently (Mendels Second Law), look at each
gene pair separately. For Aa x Aa, the F1 would show a 3 [AA, Aa, Aa
or, more simply, A-] : 1 [aa] phenotypic ratio [i.e., 3 A- / 1 aa].
o Within the of F1 showing the A phenotype [genotype A-], the B
gene segregates 3:1 [3/4 B-, 1/4 bb]
o Within the of F1 showing the a phenotype [genotype aa], the B gene
segregates 3:1 [3/4 B-, 1/4 bb]

In the cross to the left,

purple (W) is dominant
over white (w), and tall
(S) is dominant over short
(s). The cross is WwSs x
WwSs
Because the genes for
color and size assort
independently (i.e., are
independent events), the
multiplicative rules of
probability can be applied
to predict the ratio
offspring showing various
phenotypes.

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Application of multiplicative rules of probability for a tetrahybrid cross

in which all genes assort independently from each other.
The cross: TtGgCcSs x TtGgCcSs

try doing that with a Punnett square [and finishing the test on time

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Analysis of predictions of Mendels Second Law using a Testcross

o What is a testcross?
o Definition of a tester strain
o What is the value of a testcross in testing Mendels Second Law?

Testcross of a double heterozygote.

If the dihybrid was CcSS,
how would that alter the
result of the testcross?
CCSS? CCSs?

While were at it

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What is a backcross?
What is it used for?

23

Backcrosses are used when one desires to introduce a specific trait into
a particular parental genetic background [i.e., I want to introduce a
different allele into a parental background, but I want the resulting
strain to be as identical to a parental genetic background as possible]

Each successive backcross to the original parent [the elite strain in the
above Figure] increases the gene complement of that parent in the resulting
progeny.

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Introducing a desired recessive trait into a particular genetic background by

backcrossing is a touch more complicated..

Note: Blue arrows indicated selfing crosses. [Ques: Why a selfing cross?
Why not cross it with the original rr strain from the P generation?]

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Analyzing large numbers of progeny is great for statistical significance, but

the data typically do not fit exactly a specific ratio. How does one
determine if your data is consistent with a particular ratio?
P

RR yy
x
rrYY
(round, green) (wrinkled, yellow)

F1

RrYy
(round, yellow)

F2

315 round, yellow

101 wrinkled, yellow
108 round, green
32 wrinkled, green

Hypothesis: the data above are consistent with a 9:3:3:1 ratio [Assumption:
equal viability of all progenystated as a condition of the analysis below]
Chi square: Probability of observing a deviation from the expected results
at least as large on the basis of chance if the hypothesis.
2 = [(O E)2/E]
For the data above:
Perfect 9:3:3:1 ratio for 556 total progeny is 313:104:104:35
Therefore, 2 = 4/313 + 9/104 + 16/104 + 9/35
= 0.013 + 0.087 + 0.154 + 0.258 = 0.512
Thus, there is a better
than 90% probability of
observing a deviation
from the expected
results at least as large
on the basis of
chanceand we can
accept out hypothesis.

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The garden pea was Mendels model system. As weve already noted, other
model systems for genetic studies include bakers yeast (Saccharomyces
cerevisiae), fruit fly (Drosophila melanogaster), maize (Zea mays)
nematode (Caenorhabditis elegans), and zebrafish (Danio rerio),
BUT.
.what if we need to work in a system where we:
1. Cannot obtain a large number of offspring in each generation
2. The generation time of the investigator approximates the generation
time of the genetic system
3. You cannot make controlled crosses
for example, human genetics!
In those cases, we often must resort to pedigree analysis..retrospective
genetic analysis would be a good description.
Symbols Used in Pedigree Analysis

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Some Examples..

Autosomal Recessive Trait

For rare traits, most affected individuals are the children of unaffected parents. For
extremely rare traits, the unaffected parents may be related to each other
The risk of an affected child from mating of two heterozygotes is 25%
Males and females are affected in roughly equal proportions
All children of two affected individuals are affected
Trait often seen to skip generations

Q. Can you determine the genotype for each of the individuals in the pedigree above?
[Note: it may not be possible to infer the exact genotype in each case]

Consanguineous matings have a nasty habit of revealing the presence of

(rare) recessive traits within a family..

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Autosomal dominant trait

All affected individuals have at least one affected parent (exception in cases where
gene has high mutation rate..i.e., sudden appearance of an inheritable allele, not
transmitted via either parent)
Males and females affected in roughly equal proportions.
The phenotype is often more severe in affected homozygotes
Two affected individuals will have unaffected offspring if the parents are both
heterozygotes
Trait generally does not skip generations (unless penetrance is reduced)

X-linked dominant trait

o
o
o
o

Affected females are generally heterozygous

If an affected male has an affected son, then the trait is not X-linked (Why??)
All daughters of affected males are affected; all sons are unaffected
A heterozygous (affected) female will transmit trait to half of her offspring (males
and females equally affected)

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X-linked recessive trait

Much more common in males than females

Hemizygous males and homozygous females affected
Sons of heterozygous females have 50% chance of being affected (mother is not
affected)
Sons of affected females always affected (unless penetrance is reduced)
Affected males transmit mutant allele to all of their daughters, but none of their sons.
Affected males produce unaffected daughters that, in turn, produce affected sons
[some refer to this as skipping a generation].

Y-linked inheritance

Appears only in males

Aassed directly from father to son
About two dozen traits known

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Mitochondrial Inheritance [a type of non-Mendelian inheritance]

o Maternal mode of inheritance [i.e., affected males produce normal offspring;

affected females produce only affected offspring]
o Hereditary mitochondrial diseases usually affect tissues with high needs for
energy income as neuromuscular or central nervous system. Common
symptoms of mitochondrial diseases are encephalopathy, ataxia, spasms,
(cardio)myopathy, deafness or diabetes mellitus.
Inheritance of genetic disease/traits in humans (p = short arm; q = long arm)
.. Want more? Go here: http://ghr.nlm.nih.gov/
Autosomal recessive
Cystic fibrosis (CFTR gene; 7q31.2); Gauchers disease [GBA or -glucocerebrodase
gene; 1q21); Xeroderma pigmentosum [ERCC2, ERCC3, POLH, XPA and XPC genes,
many involved in NER or nucleotide excision repair.i.e., repair of DNA damage]
Autosomal dominant
Huntingdons disease (huntingtin or HTT gene; 4p16.3), Familial hypercholesterolemia
(LDLR gene; 19p13.2); Marfan syndrome (FBN1 or fibrillin 1 gene; 15q21.1)
X-linked recessive
-X-linked adrenoleukodystrophy [Addisons disease is one manifestation; ABCD1 gene,
or ATP-binding cassette, subfamily D; Xq28; causes defects in processing of very long
chain fatty acids; heterozygous females can show some symptoms..i.e., some
mutations may show partial dominance..not an uncommon phenomenon!!]
-Menkes syndrome [copper transport disease; ATP7A gene or ATPase, Cu++transporting, alpha polypeptide; Xq21.1]
X-linked dominant
X-linked hypophosphatemic rickets [PHEX gene, or phosphate-regulating endopeptidase
homolog, X-linked; Xp22p2-p22.1; this is the most common hereditary form of the
disease; there are other formsX-linked recessive, autosomalthese are due to
mutations in other genes that have a similar effect on phosphate levels as PHEX]
Mitochondrial
Leber hereditary optic neuropathy (LHON) [mutations in MT-ND1, MT-ND4, MT-ND4L,
or MT-ND6 ; all affect the function of complex I (NADH dehydrogenase complex)
involved in electron transport]

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