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DNA Technology Techniques and Applications

This document discusses recombinant DNA technology and genetic engineering techniques. It provides examples of using DNA analysis to solve criminal cases by identifying DNA evidence and matching samples to suspects. It also describes producing human insulin through genetic engineering and using polymerase chain reaction to amplify DNA.

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Ali Al-hashemi
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© All Rights Reserved
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0% found this document useful (0 votes)
247 views8 pages

DNA Technology Techniques and Applications

This document discusses recombinant DNA technology and genetic engineering techniques. It provides examples of using DNA analysis to solve criminal cases by identifying DNA evidence and matching samples to suspects. It also describes producing human insulin through genetic engineering and using polymerase chain reaction to amplify DNA.

Uploaded by

Ali Al-hashemi
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

21 Recombinant DNA technology

AQA Biology Exam-style questions

1 The diagram below shows the results of a test which can be used to analyse
evidence left at the scene of a crime. This can then be compared with samples
taken from various suspects.

a i Name this technique.

(1 mark)

ii State two types of evidence left at the scene of the crime which may have
been used to provide the DNA sample.

(1 mark)

iii Identify the suspect incriminated by the results. Give a reason for your
answer.

(2 marks)

b Briefly describe the part played by the following in the technique:


i restriction endonuclease;

(1 mark)

ii gel electrophoresis.

(1 mark)

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21 Recombinant DNA technology
AQA Biology Exam-style questions

c Other than criminal investigations, give another situation where this


technique is used.

(1 mark)

2 Scientists manufactured large quantities of human insulin using genetic


engineering.
They started by isolating mRNA from pancreas cells. From this they produced
DNA which coded for insulin.
a i Suggest two reasons why it was better to start with mRNA from pancreas
cells rather than with the DNA from these cells.

(2 marks)

ii The scientists used two enzymes, Enzyme 1 and Enzyme 2, to produce


DNA from mRNA.
The reactions catalysed by these enzymes are shown below.
Enzyme 1
mRNA single-stranded DNA
Enzyme 2
single-stranded DNA double-stranded DNA
Name enzymes 1 and 2.

(2 marks)

iii In a double-stranded DNA molecule, the two strands are held together by
weak bonds.
Name this type of bond.

(1 mark)

The scientists used the polymerase chain reaction (PCR) to make copies of the
DNA.

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21 Recombinant DNA technology
AQA Biology Exam-style questions

The diagram shows the stages of the PCR.

b i P and Q are short lengths of single-stranded DNA.


What name is given to molecules such as P and Q?

(1 mark)

ii The mixture is cooled from 95 °C to 55 °C at step 2.


Explain why.

(1 mark)

iii Explain the function of molecules P and Q.

(2 marks)

iv How many copies of each original DNA molecule would be present after
five cycles of PCR?

(1 mark)

AQA Biology, 2011, Unit 4 (Question 9)

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21 Recombinant DNA technology
AQA Biology Exam-style questions

3 a Scientists can use restriction enzymes to cut a sample of DNA into shorter
pieces. The lengths of the DNA pieces can be measured in units called
‘base pairs’. Explain why a ‘base pair’ is a suitable unit for measuring the
length of a piece of DNA.

(2 marks)

b Sickle-cell anaemia is caused by a point mutation in the haemoglobin gene.


Doctors have developed a test for the allele causing sickle-cell anaemia
using a restriction enzyme called Ddel. The allele for normal haemoglobin,
HA, can be cut by Ddel. The allele for sickle-cell haemoglobin, HS, cannot be
cut by Ddel.
The method used by the doctors involves the following steps.
1 Take samples of cells from a person and extract the DNA.
2 Treat the DNA with restriction enzymes other than Ddel to cut the DNA
into many fragments.
3 Carry out PCR (the polymerase chain reaction) using primers that are
specific for part of the haemoglobin gene.
4 Add Ddel enzyme to the DNA produced by the PCR.
5 Find the lengths of the resulting DNA pieces.
i How can a primer be specific to part of the haemoglobin gene (step 3)?

(1 mark)

ii Why is it necessary to carry out PCR on the extracted DNA (step 3)?

(1 mark)

c Mustapha and Shahira are both carriers of sickle-cell anaemia. Shahira is


pregnant and they want to know if their baby will have sickle-cell anaemia.
The doctors tested samples of DNA from Mustapha, from Shahira and from
the foetus. They used the method described in part b to test for the allele
responsible for sickle-cell anaemia. The primers used in this method were
specific for the base sequences that occur 55 base pairs before the point of
mutation and 55 base pairs after the point of mutation in the haemoglobin
gene.
The table shows the results.
Length of DNA pieces
Source of DNA after digesting with
Ddel/base pairs
Mustapha 110 and 55
Shahira 110 and 55
Foetus 110 only

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21 Recombinant DNA technology
AQA Biology Exam-style questions

i Explain the results of the tests on Mustapha’s and Shahira’s DNA.

(3 marks)

ii What can the doctors tell Mustapha and Shahira about the risk of their
baby having sickle-cell anaemia? Explain your answer.

(2 marks)

AQA Biology, 2012, Unit 4 (Question 8)

4 ‘Bt maize’ is a crop plant that contains the Bt gene from a bacterium. This gene
produces a protein that poisons insects that eat the maize.
Scientists used a bacterium called Agrobacterium to get the Bt gene into the
maize.
This bacterium contained a plasmid with the Bt gene and a gene for resistance
to a substance called kanamycin.
The diagram shows stages in the transfer of the Bt gene into the maize.
Note: Question 4 continues on the next page.

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21 Recombinant DNA technology
AQA Biology Exam-style questions

a The growth medium containing kanamycin (stage 3) is used to select for


maize tissue that contains the Bt gene. Explain how.

(1 mark)

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b All the cells of the maize plants produced at stage 4 contain the Bt gene.
Explain why.

(2 marks)

c Kanamycin is an antibiotic. Some scientists are concerned about the gene for
resistance to kanamycin being present in maize which is eaten by humans.
Suggest why.

(2 marks)

d The caterpillar of the European corn borer moth feeds on maize plants. When
the corn borer caterpillar eats Bt maize, the Bt protein is activated by the
unusually high pH (pH10) in the insect’s mid-gut. This activated protein
attaches to a specific receptor protein on the cell membranes of cells lining
the insect’s gut. This causes the gut cells to die and kills the insect. Scientists
have demonstrated that the Bt protein is not poisonous to humans and that it
is safe for humans to eat Bt maize.
Use this information to suggest why the Bt protein is not poisonous to
humans.

(3 marks)

AQA Biology, 2012, Unit 4 (Question 6)

5 Scientists wanted to measure how much mRNA was transcribed from allele A of
a gene in a sample of cells. This gene exists in two forms, A and a.
The scientists isolated mRNA from the cells. They added an enzyme to mRNA to
produce cDNA.
The scientists used the polymerase chain reaction (PCR) to produce copies of
the cDNA. They added a DNA probe for allele A to the cDNA copies. This DNA
probe had a dye attached to it. This dye glows with a green light only when the
DNA probe is attached to its target cDNA.
a Explain why this DNA probe will only detect allele A.

(2 marks)

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21 Recombinant DNA technology
AQA Biology Exam-style questions

b The scientists used this method with cells from two people, H and G.
One person was homozygous, AA, and the other was heterozygous, Aa.
The scientists used the PCR and the DNA probe specific for allele A on the
cDNA from both people.
Figure 1 shows the scientists’ results.

Figure 1
i Explain the curve for person H.

(3 marks)

ii Which person, H or G, was heterozygous, Aa? Explain your answer.

(2 marks)

AQA Biology, 2014, Unit 5 (Question 8)

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Common questions

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Agrobacterium transfers the Bt gene into maize cells through a plasmid vector that also contains a kanamycin resistance gene . When maize cells are grown on a medium containing kanamycin, only those that have successfully incorporated the plasmid (and thus the Bt gene) survive, as they express kanamycin resistance, acting as a marker .

Starting with mRNA allows researchers to obtain a transcript that directly represents the coding sequence of the gene without introns, facilitating the synthesis of a functional complementary DNA (cDNA). This approach avoids the complex regulation and splicing issues associated with genomic DNA .

The technique is called DNA profiling or DNA fingerprinting. Restriction endonucleases cut DNA at specific sequences, producing fragments of varying lengths that are unique to individuals . Gel electrophoresis then separates these DNA fragments by size, creating a banding pattern that can be compared to DNA samples from suspect and crime scene evidence .

Mustapha and Shahira have one band at 110 bp and another at 55 bp, indicating they are both carriers with heterozygous genotypes (HS/HA). Since their foetus's DNA showed only a 110 bp band, it indicates homozygosity for the sickle-cell allele (HS/HS), meaning the child will have sickle-cell anemia .

The PCR results will show a single intense signal for homozygous AA individuals, as both alleles bind the probe, increasing signal intensity . For heterozygous Aa individuals, the signal intensity will be reduced since only half of the alleles (the A alleles) can bind the probe, resulting in a distinct curve pattern when compared to the AA individual .

The Bt protein is activated at the high pH found in the insect gut (pH 10), where it binds to specific receptors on gut cells, leading to cell lysis and insect death . Humans have much lower pH in their digestive systems, which do not activate the protein, and lack the specific gut cell receptors, making it non-toxic to humans .

The cooling step to 55 °C allows primers to anneal to their complementary sequences on the single-stranded DNA template . Primers are short DNA sequences that initiate DNA synthesis, providing a starting point for DNA polymerase to extend and replicate the target DNA region .

Restriction enzymes like Ddel can distinguish between alleles by cutting DNA at specific sites only present in the normal allele . PCR amplifies the hemoglobin gene region, allowing the detection of whether the enzyme has cleaved the DNA. If the DNA is not cleaved (as with the sickle-cell allele), the resulting fragment length can identify the presence of the mutant allele .

A DNA probe is complementary to a unique sequence present only in allele A. The probe binds specifically to its target sequence due to complementary base pairing, ensuring that only allele A is detected, highlighted by the attached dye . This specificity is crucial for accurate detection and differentiation between alleles .

The incorporation of antibiotic resistance genes into food crops raises concerns about potential gene transfer to pathogenic bacteria, which could exacerbate antibiotic resistance issues in clinical settings . This risk could undermine the efficacy of antibiotics used to treat human infections, posing significant public health challenges .

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