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Anderson Junior College

JC2 9744 H2 Biology


Term 2 Block Test Answers
Modified NYJC Prelim 2016 P3 SQ2

1 A violent air crash resulted in the death of 150 passengers. The poor condition of the crash
victims made physical identification impossible. Forensic scientists were tasked to confirm the
identities of victims by using molecular techniques for families who have come forward to claim
the correct remains of their relatives.
DNA samples of human remains and surviving relatives were subjected to restriction enzyme
digest and Southern blot analysis to analyse band patterns. Single locus probes will yield DNA
patterns of seven individuals at a single locus, as shown in Fig. 2.1. On the other hand, multi-
locus probes can yield more information about each individual since 10 to 30 loci can be
simultaneously analysed. This is illustrated in five different individuals in Fig. 2.2.

Individual

Individual

Fig. 2.1
Fig. 2.2
(a) (i) Describe Southern Blot technique after the gel is treated with alkali to denature DNA.
1. Transferred over to nitrocellulose membrane via capillary action;
(And bake it at 80°C to bind the DNA to the membrane)

2. Hybridize/ anneal target DNA to radioactive (single-stranded) probe;

3. via complementary base pairing by forming hydrogen bonds between single-


stranded DNA & single-stranded probe;
(Wash off excess probe);

4. Visualize dark bands with autoradiography by (describing) placing


photographic/X ray film over the nitrocellulose membrane;
Examiner’s comments:
 This question was repeated from lecture test. If you did not get full marks, it is because
you have missed out the keywords (bold and underlined).
 It is nitrocellulose membrane, not nitrocellulose paper. I did not penalise for now, but I
will for prelim.
 Baking at 800C is NOT the reason why probe binds to target DNA.

(ii) With reference to Fig. 2.1, explain the different numbers of fragments seen in different
individuals.
1 For individuals with 2 bands, they are heterozygous for that gene/ have 2 different
alleles;
2 For individuals with 1 band, they are homozygous for that gene/have the same allele.
Additional marking point:
1 For individuals with 2 bands, there are 3 restriction sites where the restriction enzymes
cut to yield different sized restriction fragments/ Individuals who have single band only
have 2 restriction sites. Therefore only 1 thicker DNA band is produced;
Examiner’s comments:
This question was well done by those who understood the question.
(iii) With reference to Fig. 2.1 and Fig. 2.2, suggest why using multi-locus probes is
preferred.

1 [Reference to Fig.2.1]: Individual 2 and 7 are indistinguishable if single probe is used.


But if multi-probe is used, they can be distinguished.
2 [Explanation]: This is due to more bands/ more unique patterns/ more basis for
comparison.
Examiner’s comments:
Only a small handful made reference to individual 2 and 7.
(b) DNA can be analysed by Southern blotting or by PCR analysis.
Describe two differences between Southern blotting and PCR analysis in the analysis of
the single locus VNTR shown in Fig. 2.1.

Southern blot analysis PCR

Single stranded DNA/RNA radioactively (Forward and reverse) primers that flank
labelled probes hybridize/anneal to the target sequence anneal and amplify;;
target DNA sequence for detection via
autoradiography;
Feature – Use of short chemically
synthesized DNA strands.

Probe used can be s.s. DNA/ RNA Primers used must be s.s. DNA
Feature – nature of nucleic acid used

Restriction enzymes are used to cut Taq DNA polymerase (catalyses


target DNA from genomic DNA at formation of phosphodiester bonds)
recognition;; between adjacent nucleotides
Feature – methods to get target DNA

Band of interest is probed with denatured Size of PCR products can be analysed/
radioactively-labelled DNA sequence for visualised directly using ethidium
visualisation by autoradiography;; bromide/ methylene blue;;
Feature – methods to visualize

DNA is denatured by alkali DNA in PCR is denatured by heat.


Feature – methods to denature DNA

[Total: 10]
ACJC Prelim 2014 P2 SQ4
2 A group of genetics students were examining the inheritance of fruit colour and shape in a
particular species of plant. When they crossed a pure breeding plant that had red, oval fruits
with pure breeding plants that had yellow, long fruits, all the offspring produced were observed
to have red, oval fruits.

They then proceeded to testcross two plants, A and B, from the F2 progeny, both having red,
oval fruits and the following results were obtained as shown in Table 4.1.

Table 4.1

Phenotype Progeny of A Progeny of B

Red, long fruits 46 4

Yellow, oval fruits 44 6


Red, oval fruits 5 43
Yellow, long fruits 5 47
(a) Based on the information and results of the crosses given above, describe the inheritance of
fruit colour and shape in this species of plant.

1. Two genes located on the same chromosome/linked;


2. Red allele dominant to yellow allele, and oval dominant to long fruits;
@ 1m
[2]

Examiners’ comments:
 Most candidates could tell that this is a linked genes question.
 Candidates have to state that it is the allele that are dominant and traits.
 Most common mistake is using the term “gene” when it should be “allele”. Answers such as
“the gene coding for the red colour…” is wrong. The gene is fruit colour, encoded for by 2
different alleles, red and yellow. The 2 terms cannot be used interchangeably.
 Answer such as “dominant allele codes for red and oval fruit” is also not accepted. One allele
cannot code for both phenotype. Language is important.

(b) Using appropriate letters to represent the various alleles, state the genotypes of plant A and B.

Letter to represent allele for red fruit : R


Any appropriate letters and
Letter to represent allele for yellow fruit : r both correct @ 1m

Letter to represent allele for oval fruit : A [1]

Letter to represent allele for long fruit : a

Genotype of plant A:
(with red, oval fruit)

Genotype of plant B:
(with red, oval fruit)

Examiners’ Comments:
 Since this is a linked gene question, genotype representation has to be using the “stick
figure” so that it is possible to check if the correct alleles are linked together on the
chromosomes.
 Plant A and B are from the F2 generation selfed from F1. Since both plant A and B shows all
4 phenotypes, they must both be heterozygous for fruit colour and shape gene. The higher
number of offspring indicates the parental phenotypes and an indication of which alleles are
linked together.
 For plant A: In a test cross, the homozygous recessive parent contributes (ra) allele. For
parental phenotype to be red and long (no crossing over occurs between the 2 gene loci),
the parental gamete for A has to be (Ra) to produce a red, long phenotype. Since Plant A is
a heterozygote, this means the other chromosome is linked (rA).
(b) Draw a genetic diagram to explain the results of the testcross on plant A as shown in Table 4.1.
Phenotype of parents: Red, oval fruits x yellow, long fruits

Genotype of parents: x ;

Gametes: (Ra) (rA) (RA) (ra) x (ra) (ra) ;

F1 genotypes: : : : ;

F1 phenotypes: 46 Red, long : 44 Yellow, oval : 5 Red, oval : 5 Yellow, long ;

Majority, from gametes Minority, recombinant


with no crossing over; offspring;
Parental phenotypes Recombinant
phenotypes

 1 m for correct genotypes of parents with cross in between


 1 m for correct gametes with cross in between
 1 m for correct F1 genotypes
 1 m for correct corresponding phenotypes
 1 m for annotations on recombinants (either at the gametes or phenotype level)
[5]

Examiners’ comments:
 Many students experienced difficulty in this question if they couldn’t figure how the alleles
are linked in the previous question.
 Some students again, did not use the “stick figure” representation at the parental genotype
level and hence, couldn’t check for the correct alleles on the same chromosome. This makes
it impossible to check if the gametes are correct and therefore no marks would be awarded.
 Students need to be sensitive to wrong answers. If your genetic diagram shows there are
more recombinant phenotypes than parental phenotypes, this is a clear sign that the alleles
are not represented correctly.
 There are a significant group of students that are still unable to use the correct format of
genetic diagram, with missing labels, titles, did not quote phenotypic numbers or are
incomplete.
The group of students proceeded to measure the fruit length obtained from true breeding plants with
oval fruits and those from true breeding plants with long fruits. Table 4.2 below shows the number of
fruits in each fruit length category from the two respective pure breeding plants.

Table 4.2

Fruit length / cm
3 4 5 6 10 11 12 13
Plants with
4 7 6 2
oval fruits
Plants with
3 6 9 5
long fruits

(c) Explain the variations observed in fruit length within each group of pure breeding plants.

1. Variation in fruit length due to environmental factors (such as amount of sunlight (


for PS) / location of fruit on plant, number of fruits on same branch / availability of
water / nutrients during development);

2. Pure breeding plants are homozygous at all gene loci;

@ 1m
[2]
[Total: 10]
Examiners’ comment:
 Most students could identify that fruit length shows continuous variation. Only a handful of
students mistakenly viewed this as discontinuous variation.
 However, only the more able students could state that since the plants are pure breeding, they
are homozygous at all gene loci. Therefore, any variation seen within the group cannot be
attributed to genetic contribution. Answers relating to additive effect of genes, each gene having
a small effect are not relevant for this question.
 Therefore, within each group, variation is due to environmental factors.
MJC Prelim 2015 P2 SQ5 (modified)

3 The mode of inheritance for shape of summer squash can be investigated by a cross between two
summer squash plants with disc-shaped fruit. The cross resulted in offspring with 3 different
phenotypes in the following numbers:

Disc-shaped fruit 1071


Sphere-shaped fruit 720
Long fruit 123

Shape of summer squash is determined by 2 gene loci, A/a and B/b.

a) Explain how different fruit shapes in summer squashes come about.

 The fruit shapes in summer squashes are controlled by epistasis, where there is duplicate
genes with cumulative effect. [No need to indicate recessive/dominant epistasis]

 Presence of a dominant allele from either gene / A and B allele each codes for an enzyme
 If dominant alleles from both genes are present / genotype A_B_, the fruit becomes disc-
shape / A_bb and aaB_ produce sphere plants/ Absence of any dominant alleles
homozygous recessive plant aabb, and hence resulted in long fruits.
Examiner’s comments:
 Some students do not realise it is epistasis. Genotypes are also not referenced.
b) With this understanding of their mode of inheritance, a breeder crossed two sphere-shaped
summer squash plant together. None of the offspring produced gives sphere-shaped fruit.

Construct a genetic diagram to explain the cross.

Parental phenotype Sphere-shaped x Sphere-shaped

Parental genotype AAbb x aaBB


[1m – correct parental genotypes]

Gametes Ab x aB

F1 genotype AaBb

F1 phenotype All Disc-shaped fruit

[1m – correct F1 genotypes AND phenotypic ratio]

Examiner’s comments:
Generally well done
c) A test cross is used to determine the genotype of an organism. However, a test cross may not
be able to determine the genotype of a disc-shaped squash entirely. Explain why.

 Test cross means cross the disc-shaped squash plant with a long-shaped squash plant that
is homozygous recessive / aabb;

 Not able to differentiate between plant that is heterozygous at either one loci / AaBB or
AABb, the offspring will have the phenotype of disc-shaped fruit and sphere-shaped fruit;

A test cross is only effective in determining the genotype when:


 the disc-shaped squash plant is homozygous at both gene loci / AABB, all the offspring
will have disc-shaped fruit; OR
 the disc-shaped squash plant is heterozygous at both loci / AaBb, the offspring will have
all 3 phenotypes;
Any 2

Examiners’ comments:
 Question proved challenging for most students.
 Some students vaguely explained that because there are many genotypes that can result in
 a disc shaped squash, it is not possible to determine genotype entirely. That is not true.
 A number of students incorrectly stated that aaB_ would result in a disc shape squash.
 A test cross allows us to determine unknown genotype of parent by observing offspring
phenotypic ratio. If a test cross is not able to determine the genotype, this means that there
are certain genotypes that will produce the same phenotypic ratio, therefore not
discriminating.
 Some students mentioned that if sphere shaped offspring are produced, we are unable to
differentiate if the genotype of parental disc shaped squash is AaBB or AABb. This is not
accurate as AaBb parental squash will also generate sphere shaped offspring. Therefore, it
is important to state that offspring produced is 1 disc: 1 sphere shaped offspring.

Squashes also possess different fruit colours. Fruit colours of squash are determined by two other
gene loci on 2 different chromosomes. The colours of squash are determined by gene locus C/c.
The presence of a dominant allele, C results in white squash, while the recessive allele gives rise to
coloured squash. At the second gene locus G/g, allele that codes for yellow is dominant to allele
that codes for green.

A cross between two heterozygous white squash plants gives phenotypes in the following numbers:
White squash 250
Yellow squash 60
Green squash 10

The expected ratio for the above cross is 12:3:1.

The 2 distribution table and equation to calculate 2 is shown below.

number of degrees probability


of freedom (v) 0.05 (𝑂− 𝐸)2
1 3.84  2
=Σ 𝐸
2 5.99
3 7.82
4 9.49
d) Using the equation and 2 distribution table given above, explain if epistasis is the correct
explanation for the results obtained.

(240−250)2 (60 −60)2 (10−20)2


2 = + + = 0.42 + 0 + 5 = 5.42
250 60 20
 Calculated  = 5.42.
2

 At degree of freedom = 2, calculated 2 is less than the critical value of 5.99 OR


The probability that the difference between expected and observed number is due to
chance is more than 0.05 / p >0.05.
 Difference is not significant / due to chance, the observed number conform to the
epistatic ratio 12:3:1, hence epistasis is the correct explanation.

Examiners’ comments:

 A significant number of students are not able to calculate chi-square correctly.


 Observed and expected numbers of offspring should be used to calculate chi-square. Not
ratio.
 Some students gave contradictory answers such as calculated chi-square is less than critical
chi-square, probability that difference is due to chance is less than 0.05. Such contradictory
answers cannot be awarded marks.
 A handful of students stated that there is a “high probability” that difference is due to chance.
Always refer to probability 0.05. It should be “probability that difference is due to chance is
more than 0.05”.
 Students who could remember structure of answering such chi-square questions did well.

[Total: 10]
Essay

Explain how the environment may affect the phenotype. [10]

Comments:
Key to success in such essay questions is to quote many examples. You’ll need about four. Do not
overwrite on only one or two examples as you’ll only be able to get 6 marks max.

Answers to this question can be far and wide. It can come from other topics too but the link between
environment and phenotype needs to be clear. For example, many students only state that in
plants, light induces chlorophyll synthesis. It is more accurate to extend the answers to therefore
plants will look brighter green (phenotype).

Also, a number of students used common examples eg. When there are more water and nutrients in
the soil, the plants will grow taller. While the answers are not wrong, the answers need to be
substantiated with key terms from the syllabus. Minimally, students need to link to photosynthesis.

There is no other way to ace such questions besides memorization. Some students’ answers were
vague and unclear due to lack of specific details. Pls memorize the content.

This Q is a LO in Genetics and Inheritance (Mr Poh’s lectures). The syllabus specifically states
using examples of Himalayan rabbits and honey bees. So these two examples are compulsory.
Students who did not mention these two examples will not be able to score full marks.

Many students also explained in terms of natural selection. Here, while it was a good attempt, you
need to be careful not to perpetuate the misconception that the environment will induce specific
mutations to give rise to alleles that will confer a selective advantage to the organism in order for the
organism to adapt to the environment. For example, a student wrote that in the North Pole, the cold
environment induce mutations in the polar bears to develop white fur so that the bears are
camouflaged against the environment. This is a MAJOR misconception as genetic variation is a pre-
requisite of natural selection. GV should already exist in the gene pool. The correct way is to explain
it via Natural selection with particular focus on environment and phenotype. Environment serves as
a selection pressure. Organisms with traits which enable them to adapt better to the environment
are at a selective advantage and they will be selected for. They are more likely to survive and
reproduce to pass on the alleles coding for advantageous traits to the offspring / next generation.
Over time and many generations, there will be a higher frequency of individuals with the
trait/phenotype that has been selected for. This last sentence is essential as it is the link to how
environment affects phenotype.

Answers on discontinuous variation is rejected as the environment only has a small effect.

Another point to note is that high ability students addressed the question in terms of the genes as
well. For example, late onset of diabetes develops in genetically diabetic individuals who overeat
but not in genetically diabetic individuals whose diet is low in sugar and lipids. The idea is that
environment can lead to a different phenotype that was coded for by the genes.

Organisation of answers is important. You should use headings.

1. Continuous variation is due to action of polygenic inheritance (three or more genes)


and the effects of these genes are additive
2. Effect of individual genes cannot be observed/ Each gene contributes to a small effect
on the phenotype and therefore the environment has a large effect on the phenotype.

Natural Selection
1. Challenges in the environment (e.g. climate, disease, competition from other
organisms, predation and limitation of food and other resources) will exert selection
pressure on the individuals of the population.
2. Organisms with traits which enable them to adapt better to the environment are at a
selective advantage and they will be selected for. They are more likely to survive
and reproduce to pass on the alleles coding for advantageous traits to the offspring /
next generation.
3. Over time and many generations, there will be a higher frequency of individuals with the
trait/phenotype that has been selected for.

 Honey bees (As Specified in the Syllabus)

1 Queen and workers have same amount of genetic material but are phenotypically
different.
2 Workers are sterile, smaller, have larger mouthparts & modified legs.
3 Larvae destined to be workers are fed honey and pollen on the third day onwards
whereas those destined to be queen continue with royal jelly.
4 The high protein of royal jelly stimulates the formation and maturation of the female
reproductive system

 Late onset of diabetes

1. Develops in genetically diabetic individuals who overeat but not in genetically diabetic
individuals whose diet is low in sugar and lipids.
2. Overeating of sugary foods for a long period of time causes repeated stimulation of
the pancreas leading to prolonged secretion of insulin.
3. Repeated exposure of target cells to large amounts of insulin desensitizes the cells’
responsiveness to insulin.
4. This may result in target cells failing to reduce high blood glucose level in the usual
way, resulting in diabetes.

 Phenylketonuria (PKU) in humans

1. PKU is a recessive disorder of amino acid metabolism


2. Infants homozygous for the mutant allele accumulate toxic substances in their brains
due to having no functional Phenylalanine Hydroxylase (PAH) enzyme.
3. These substances are not lethal but can impair mental ability by affecting the brain’s
development.
4. A particular amino acid, Phenylalanine is a precursor to the toxic substance that
accumulates in human brain. This amino acid is ingested in the diet.
5. Infants with PKU who are fed normal diet ingest enough phenylalanine to bring out the
worst manifestations of the disease/ Infants who are fed low phenylalanine diet usually
mature without serious mental impairment.

 Effect of temperature on phenotype - Vestigial wings in Drosophila

1. In Drosophila, the allele for vestigial wing is recessive to that for long wing.
2. Expression of vestigial wings is affected by the temperature at which the insect
develops.
3. Allele for vestigial wings is expressed only at low temperatures.
4. Drosophila flies that are homozygous recessive for vestigial wings, will develop the
phenotype at 21oC, intermediate wings at 26oC and long wings at 31oC.

 Coat colour in Himalayan rabbits (As Specified in the Syllabus)

1. The Himalayan Rabbit has a white body with black fur on its extremities/ black ears,
nose, feet and tail.
2. Black fur absorbs light and solar heat, keeping that part of the rabbit which has dark fur
warmer/ The extremities are coldest due to less blood flow.
3. If the fur on the back is shaved, an ice pack is fixed onto the rabbit’s back and left in
position for weeks (kept cold constantly), Black fur begins to develop beneath the ice
pack.
4. This is because the enzyme tyrosinase that makes the pigment, is active at lower
temperature but is denatured at higher temperatures.