1 Fig. 22 shows four nucleotides.

Fig. 22

(i) On Fig. 22, use the letter R to label a bond that would be made by the action of a ligase enzyme.

[1]

(ii) On Fig. 22, use the letter P to label a bond that would be broken during the hottest step of the PCR
reaction.

[1]

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 2 *Scientists have genetically engineered many different species of plant.

Aubergine plants, Solanum melongena, can suffer damage from moth larvae.

Scientists have produced a variety of aubergine that is resistant to moth larvae. To create the
resistance, scientists transferred a gene from the Bacillus thuringiensis bacterium.

Describe the process the scientists could have used to produce the pest-resistant aubergines.

[6]

3(a) Fig. 21 shows some of the steps involved in producing a genetically modified bacterium.

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 Fig. 21

The following passage describes steps A and B. Complete the passage using the most appropriate
terms.

A gene is cut from the DNA of the donor organism using a ………………………………
……………………… .
The ………………………………………… enzyme is used to cut open a small piece of bacterial DNA
so that the base sequences at the end of each piece of DNA are ……………………………… .
[3]

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 (b) Describe the events that are taking place at the step labelled C.

[3]

4 Bacteria such as E. coli can be genetically engineered for use in medical science.

An example of the genetic engineering of E. coli is shown in the diagram below.

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 (i) Complete the diagram above by writing the missing words or phrases in the boxes labelled X, Y
and Z.

Answer on the diagram [3]

(ii) Suggest why the scientists used a plasmid that contained an antibiotic resistance gene.

[1]

(iii) The scientists observed the following:

• 1 in 400 bacteria took up the plasmid

• 1 in 1000 of the plasmids taken up by bacteria contained the β-galactosidase gene.

Calculate the percentage of bacteria that contained the β-galactosidase gene.

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 percentage of bacteria = ....................................................... % [2]

(iv) A technique called quantitative PCR is used to check that the E. coli population is growing on the
mice liver tumours rather than on healthy tissue.

Suggest how the scientists could use PCR to compare E. coli growth rates on cancerous liver
tissue and healthy tissue.

[2]

(v) Some people think that the genetic engineering of certain organisms is unethical.

However, there are very few ethical concerns about the genetic engineering of bacteria such as E.
coli.

Suggest why there are very few ethical concerns about the genetic engineering of E. coli.

[1]

END OF QUESTION PAPER

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 Question Answer/Indicative content Marks Guidance

1 i R used to label a phosphodiester bond 1

✓

ii p used to label a hydrogen bond ✓ 1

Total 2

2 Please refer to the marking instructions

on page 4 of this mark scheme for
guidance on how to mark this question.

In summary:
Read through the whole answer. (Be
prepared to recognise and credit
unexpected approaches where they
show relevance.)
Using a ‘best–fit’ approach based on
the science content of the answer, first
decide which of the level descriptors,
Level 1, Level 2 or Level 3, best
describes the overall quality of the
answer.
Then, award the higher or lower mark
within the level, according to the
Communication Statement(shown in
italics):

award the higher mark where the

Communication Statement has
been met.
award the lower mark where
aspects of the Communication
Statement have been missed

The science content determines the

level.
The Communication Statement
determines the mark within a level.

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 Question Answer/Indicative content Marks Guidance

Level 3 (5–6 marks) 6 Indicative scientific points may include:

Describes the process in detail, with no
significant errors.
method for gene extraction from the
There is a well-developed line of bacterium (e.g. conversion of
reasoning, which is clear and logically- mRNA to cDNA with reverse
structured and uses scientific transcriptase, or removal of gene
terminology at an appropriate level. All with restriction enzymes)
the information presented is relevant use of appropriate vector (e.g. Ti
and forms a continuous narrative. plasmid of
Agrobacteriumtumefaciens)
Level 2 (3–4 marks) electroporation
Describes some details of the process, use of DNA ligase
with only minor errors. reference to marker genes and their
purpose
There is a line of reasoning presented electrofusion
with some structure and use of
appropriate scientific language. The
information presented is mostly
relevant.

Level 1 (1–2 marks)

Describes aspects of the process, but
with significant omissions or errors.

The information is communicated with

only a little structure. Communication is
hampered by the inappropriate use of
technical terms.

0 marks
No response or no response worthy of
credit.

Total 6

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 Question Answer/Indicative content Marks Guidance

3 a restriction, enzyme / endonuclease  3 max

same  ALLOW restriction (endonuclease)

complementary  IGNORE sticky ends

Examiner’s Comments
This was generally well answered by
most candidates. The most common
incorrect response was the third, where
several candidates put ‘sticky’ or
‘exposed’.

b the gene / the DNA fragment, inserted 3 max ALLOW the bit of DNA combines with
into plasmid  ring of bacterial DNA

complementary bases (pair / anneal) ALLOW complementary sticky ends



formation of hydrogen bonds 

formation of phosphodiester bonds 
DO NOT CREDIT in context of making
using (DNA) ligase  hydrogen bonds

Examiner’s Comments
This question differentiated well
between candidates with all marking
points seen. Common responses that
were not credited included referring to
the plasmid imprecisely as DNA, or
incorrectly as a bacterium. Many
candidates also stated that the DNA
ligase was used to form hydrogen
bonds and were not credited for
mentioning the ligase. Some
candidates described the events
occurring in step D, as opposed to C,
and gained no marks. Precise and
correct use of key terms is essential
when answering knowledge and
understanding questions such as this.

Total 6

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 Question Answer/Indicative content Marks Guidance

4 i X restriction (endonuclease) ✓ 3
Y (DNA) ligase ✓ (AO1.2)
Z electroporation / culture heating / ALLOW electric shock
heat shock / calcium salts ✓ ALLOW calcium ions

ii (acts as) marker / reporter, gene ✓ 1 max

idea of to indicate which bacteria have (AO2.5) e.g. ‘can identify transgenic bacteria’
taken up the plasmid ✓

iii 0.00025 or 2.5 x 10-4 ✓✓ 2 FIRST CHECK ON ANSWER LINE

(AO2.6) If answer = 0.00025 or 2.5 x 10-4 award
2 marks

If the answer is incorrect, award one

mark for
1/400 = 0.0025 or 2.5 x 10-3
OR
0.0025/1000 = 0.0000025 or 2,5 x 10-6
OR
0.0000025 x 100 (= 0.00025 or 2.5 x
10-4)

iv idea of extract DNA from cancerous 2 max

liver and (named) healthy tissue ✓ (AO3.4)
choose primers for, E coli / β-
galactosidase, DNA ✓
idea of comparing rate of DNA e.g. ‘compare amount of DNA after 30
amplification ✓ cycles of PCR’

v idea of safety of genetic engineering 1 max e.g. ‘It’s been done for many years
(in bacteria) has been established ✓ (AO3.2) without any problems’ / ‘genetic
engineering is safe’
idea of few animal rights issues to
consider ✓ e.g. ‘bacteria do not have emotions like
animals that can be engineered’ /
‘bacteria do not feel pain’ / ‘bacteria
are not conscious’

Examiner’s Comments

Most candidates gave appropriate

suggestions for the functions of
proteins A and C, based on the
information given in Table 4.1, and
could recognise that antibiotic A22
could cause problems in humans by

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 Question Answer/Indicative content Marks Guidance
binding to actin in muscles.

Likewise, many candidates used the

data in Table 4.2 to evaluate the
advantages and disadvantages of the
two antibiotics, gaining both marks.
However, some candidates lost marks
by not being precise in their responses,
for instance by saying oritavancin
cures fewer bacterial infections without
naming the specific infection
Streptococcus, or by saying it has
fewer side effects without naming the
side effects.

Many candidates correctly identified X

and Y in (c)(i) but few could name Z
correctly.

Few candidates knew why antibiotic

genes are used in plasmids for (c)(ii).
Many candidates referred to the gene
providing bacteria with protection from
antibiotics without stating why this was
done in this case.

Many candidates struggled to gain

both marks for the two- step calculation
in (c)(iii), although many gained a mark
for working out one of the steps
correctly.

Candidates often understood the

technique of PCR, but could not apply
this technique to compare growth rates
of E.coli in different tissues for (c)(iv).
Those that suggested taking DNA
samples from both tissues and using
PCR to compare the amounts of DNA
produced after a fixed number of
cycles gained credit here.

Total 9

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