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Gene technologies Flashcards

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AP® Biology flashcards Biology 101 flashcards
1
Q

What is the purpose of PCR

A

(Polymerase chain reaction)
to amplify quantity of DNA

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2
Q

what are the 4 items required for PCR

A

-DNA sample to be amplified
-excess of 4 nucleotide bases (to synthesise new DNA strand)
-Tag DNA polymerase- bind nucleotides together
-Primers- provide starting sequence for Tag

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3
Q

Stage 1 PCR

A

Separating DNA Strands
Temp: 90
Unzips polynucleotides by breaking H bonds between NCB

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4
Q

What is the name and function of computer used in PCR

A

Thermal cycler
-Varies temp at precise time intervals

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5
Q

Stage 2 PCR

A

Anneling Primers
Cooled to 55
Primers bond to 3’ end of DNA strands using complementary base pairing

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6
Q

Stage 3 PCR

A

Synthesis
Heated to 75 (optimum temp for Taq)
-Adds nitrogen containing bases to primers, building up complementary DNA strands
-Phosphodiester bonds form between adjacent nucleotides

-> Double stranded DNA identical to the original sequence

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7
Q

Uses of PCR

A

-Forensic science
-DNA profiling in paternity testing
-DNA amplification of DNA extracted from fossils

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8
Q

Similarities and differences between PCR and SCR

A

+free nucleotides
+ one new, one old strand in each molecule of DNA

-PCR produces only short strands (whole chro in SCR)
-PCR requires primers
-PCR requires high temps to separate DNA strands (DNA helicase in SCR)

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9
Q

Intron

A

non-coding DNA

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10
Q

extron

A

coding DNA

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11
Q

What are variable number tandem repeats

A

Patterns of repeated adjacent nucleotides

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12
Q

why does Tag DNA polymerase not denature at 90 degrees

A

it has covalent bonds and disulphide bridges

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13
Q

Use of VNTRs

A
  • genetic fingerprinting
    -inheritance matching (more distant, less relatedness)
    -Essential for forensic science
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14
Q

what is the function of electrophoresis

A

preparation and analysis of DNA for sequencing

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15
Q

How to DNA fragments separate during electrophoresis

A

-DNA fragments put into wells in aragose gel
-Buffer solution added
-Current applied
-Since DNA fragments are - (phosphate groups), they move to positive end
-Shorter fragments of DNA move faster and further

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16
Q

How can DNA bands be visualised following electrophoresis

A

Using fluorescent dye and a UV light

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17
Q

Examples of GE

A

Production of human insulin
Human growth hormones to treat dwarfism

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18
Q

What is the def of GE

A

the manipulation of an organism’s DNA

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19
Q

1 stage of GE: Gene Isolation

A

-Gene is isolated using restriction enzymes
-mRNA extracted from cells
-Reverse transcriptase produces single stranded complementary DNA (cDNA)
-DNA polymerase then produces strand complementary to cDNA
-end up with double stranded DNA

+double stranded DNA has no introns as produced from mature mRNA

20
Q

2 stage of GE: Insertion of gene into vector

A

-Vector eg. plasmid cut with same restriction enzymes
-Both plasmid and recombinant gene has complementary sticky ends
- They are annealed
-DNA ligase forms phosphodiester bonds between nucleotides of plasmid and recombinant gene-> recombinant plasmid

21
Q

3 stage of GE: Uptake of recombinant gene

A

-Put in Ca2+ solution and heat
-CSM becomes more permeable so recombinant plasmid taken up by bacteria
-Forms transgenic bacteria

22
Q

Identifying transgenic bacteria

A

-Reporter gene is also inserted into bacterium
-Recombinant gene is usually inserted into reporter gene so that it does not work
-cells not displaying reporter gene property have taken up gene

23
Q

Restriction enzymes

A

-Cuts DNA strand at specific site
-site complementary to active site of enzyme
-produce sticky ends

24
Q

What is meant by palindromic

A

sequence of bases on one strand is reversed on the other strand

25
Q

GE in cows

A

Lysozyme- antibacterial properties

26
Q

GE in goats

A

antithrombin- prevents blood clotting

27
Q

Advantages of using transgenic plants to produce human proteins

A

-low production costs
-unskilled labour sufficient to maintain crops
-minimal risks of contamination of human pathogens

28
Q

Crops have been genetically modified to:

A

-resistance to herbicides
-produce insecticides
-produce vitamin A

29
Q

How genes are inserted into GMC

A
  • T-DNA, Ti plasmid inserted into bacterium

-Gene gun- DNA coated gold particles integrates into plant chro.

30
Q

What is a knockout mouse

A

lab mouse where researchers have inactivated an existing gene by replacing it or disrupting it with an artificial piece of DNA

31
Q

why are knockout mice useful

A

provide valuable clues about what that gene normally does.
have been used to study cancer, obesity, diabetes, arthritis etc

32
Q

Disadvantages of knockout mice

A

-15% of altered embryos cannot grow into adult mice
-knocking out a gene may fail to produce an observable change

33
Q

RNA splicing

A

-pre-mrna formed at the end of transcription
-contains both introns and exons
-introns are removed
-exons are spliced together
-mature mRNA is then translated to produce ppc

34
Q

Alternative splicing

A

-some introns retained within mature mRNA and exons can be joined in different combinations
-1 gene can code for many proteins

35
Q

advantages of alternative splicing

A

increases genetic diversity

36
Q

purpose of RNA interference (iRNA)

A

control gene expression ie. can silence genes

37
Q

Small interference RNA (siRNA) vs microinteference RNA (miRNA)

A

siRNA:
must be fully complementary to mrna
more precise
double stranded

miRNA:
binds in many places in mRNA so only requires partial complimentary pairing
less precise
single stranded

38
Q

How iRNA works

A

-double-stranded iRNA binds to dicer
-dicer cuts iRNA into short sections
-short iRNA binds to argonaute protein
-1 strand of iRNA (guide strand) remains bound to argonaute protein to form RISC
-RISC complex binds to mature mRNA

39
Q

Gene therapy process

A

-adds functional therapeutic genes to cells with defective genes within them
-allows genetic disorders to be treated

40
Q

2 main problems with gene therapy

A

-getting functional genes into cell
-making functional genes work once they’re in the cell

41
Q

Severe combined immunodeficiency disease (SCID) causes

A

Faulty ADA allele
-ADA essential for proper functioning of immune system so no cell mediated response etc

42
Q

Delivery methods for functional therapeutic genes (FTG)

A

-Viruses eg. adenovirus
-Liposomes i.e. FTG wrapped in lipid molecules. Then, sprayed into nostrils, cross csm and enter cells of the lung

both ex vivo delivery methods

43
Q

Difficulties with viral vectors

A

-difficult to control where the virus inserts the gene
-if gene inserted into gene that controls cell division, it can cause cancers

44
Q

Germline gene therapy

A

adds to FTG into fertilised ovum
ensures all cells of organism will contain the FTG
so FTG can be passed on to next generation

45
Q

Why are stem cells ideal for gene therapy

A

-unspecialised
-divide by mitosis to form clones
-pluripotent, so stem cells will develop into any tissue required

46
Q

Ethical complications of germline

A

-Playing as god
-negative effects on future generations
-ethical debate- changing genomes of unborn children
-abuse of principle- designer babies?

Biology (18 decks)

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