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Biology Unit 5 > DNA Technology > Flashcards

DNA Technology Flashcards

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1
Q

How do you isolate genes

A 
Use enzymes
Either
Reverse transcriptase 
Or
Restriction endonuclease
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2
Q

How is reverse transcriptase used to isolate genes

A

Locate and extract gene
mRNA carries the code
mRNA acts as template for cDNA
cDNA formed using RT enzyme
DNA polymerase binds complementary nucleotides on cDNA template
Double stranded DNA formed using cDNA template
Copy of desired gene in DNA

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3
Q

How is restriction endonuclease used to isolate genes

A

Cuts DNA at recognition site
Cut leaves sticky ends so bigger bonding site
Complementary bases pair up
Enzyme DNA ligase joins them

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4
Q

What is In Vivo gene cloning

A

Transferring fragments to a host cell using a vector

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5
Q

What ways can DNA be inserted

A

Using plasmid vector
Using virus
Coat in lipids

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6
Q

What is the role of a vector during in Vivo gene cloning

A

Transfers genes from one organism into another

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7
Q

What vector is used to insert DNA into host cell

A

Plasmid

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8
Q

What is transformation

A

Reintroducing the plasmid back into bacterial cells

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9
Q

Describe the process of transformation

A

Mix together in medium containing calcium ions
Calcium ions and temperature change
Causes bacterial cells to become permeable
Plasmid passes through membrane into cytoplasm

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10
Q

Why do not all bacteria cells have the DNA fragment

A

Only a few take up plasmids when mixed together

Some plasmids close up before DNA fragment is incorporated

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11
Q

How do you identify which host cells have taken up the gene

A

Enzyme gene marker
Fluorescent gene marker
Replica plating using antibiotic resistance markers

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12
Q

What happens when doing antibiotic resistance markers

A

Use plasmid with 2 antibiotic resistant genes
Insert desired gene into bacteria
Do replicate plate
Put one plate with disease
If survive, shows resistant to gene which shows did not take up wanted gene
Ones we want die so know which ones on non-replica plate to keep and grow

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13
Q

How are fluorescent gene markers used

A

Jellyfish have green fluorescent protein

Transfer desired gene into plasmid from jellyfish

If bacteria cell taken up gene then they won’t fluoresce when viewed under microscope

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14
Q

How are enzyme gene markers used

A

Gene marker codes for enzyme eg lactase
Lactase turns substance from clear to blue
Desired gene placed in centre of lactase gene
If gene present, won’t cause colour to change to blue in presence of lactose

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15
Q

Why are fluorescence and enzyme gene markers more efficient than antibiotic ones

A

Won’t kill ones we want

So it’s quicker method

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16
Q

When growing bacteria, what needs to be controlled

A

Temperature

PH

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17
Q

What is In Vitro gene cloning

A

Polymerase chain reaction

18
Q

What is a thermocycler

A

Controls temperature changes

19
Q

What is polymerase chain reaction used for

A

Crime scene investigators

Info gaining about long dead organisms

Determine to identify father

20
Q

What are primers used for

A

Keep strands apart

21
Q

What is DNA polymerase used for in polymerase chain reaction

A

Join together nucleotides

22
Q

In Vitro

How are strands broke apart

A

Temperature

Break hydrogen bonds

23
Q

In Vitro

Why is heated to 95 degrees

A

To spear ate DNA strands

24
Q

In Vitro

Why is cooled to 55 degrees

A

Allowed primers to anneal

And primers to attach to start and end complementary bases on DNA strands

25
Q

In Vitro

Why is heated to 72 degrees

A

Optimum temperature for DNA polymerase

26
Q

Describe polymerase chain reaction process

A 
Put in thermocycler
Heated to 90 degrees to break bonds 
Cooled to 55 degrees
So primers can be used 
Heated to 72 degrees
Nucleotides bind to complementary bases
DNA polymerase

Now have 2 DNA molecules
Cycle takes 2 minutes
Cycle keeps going

27
Q

What are the 5 main stages of DNA technology

A 
Isolation
Insertion
Transformation
Identification
Growth/cloning
28
Q

Advantages of In Vivo

A

No risk of contamination
Very accurate
Makes copies of specific gene not whole DNA sample

29
Q

Disadvantage of In Vivo

A

Takes longer than In Vitro to produce same quantity

30
Q

Advantages of In Vitro

A

Doesn’t require living cells

Very quick process

31
Q

Disadvantage of In Vitro

A

Requires very pure sample as could contaminate with other DNA, leading to a false result

32
Q

Benefits of genetic modification

A 
Increase yield from crop
Improve nutrient content in food
Introduce resistance to disease
Making vaccines
Produce medicine for treating diseases
Develop tolerance to environmental conditions
33
Q

Describe process of Anti-thrombin production

A

Egg and sperm
Put gene in fertilised egg before division occurs
Offspring with gene are crossbred
Extracted and purified

34
Q

Benefits of recombinant DNA technology

A

Genetic fingerprinting used in forensic science
Gene therapy to cure genetic disorders
Genetically modified crop has economic and environmental advantages

35
Q

Risks of recombinant DNA technology

A

Don’t know long term effects
Gene may pass to different organisms, causes problems
What is genetically engineered gene mutates?
Expensive. Money better use eg fighting world hunger

36
Q

how can cystic fibrosis be treated using gene therapy

A

gene replacement
gene supplementation
germ-line gene therapy
somatic-cell gene therapy

37
Q

what is germ-line gene therapy

A

replacing defective gene in fertilised egg so cells of offspring will develop normally

38
Q

what 2 methods are used to deliver genes in gene therapy

A 
harmless virus (adenovirsues)
wrapping it in lipids

both then inserted in patient’s nostrils

39
Q

why are forms of delivering gene therapy not always effective

A

adenoviruses may cause infections
patient may develop immunity to adenoviruses
even if delivered successfully, few genes actually expressed

40
Q

How would you use gene therapy to treat severe combined immunodeficiency

A

Normal enzyme isolated from healthy human tissue
Then inserted into retrovirus
Retrovirus mixed with patient’s T cells
Retrovirus inject copy of normal enzyme gene into T cells
T cells reintroduced into patient’s blood, now with genetic code needed to make enzyme

41
Q

What is somatic-cell gene therapy

A

Gene therapy that targets just the affected tissue
Healthy genes are not passed on to future generations
Means treatment needs to be repeated

Biology Unit 5 (12 decks)

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