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Biology Module 2 > Gene technology > Flashcards

Gene technology Flashcards

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1
Q

What is PCR used for?

A

PCR is used to amplify target DNA sequences that are present in a DNA source

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2
Q

Describe the process of PCR

A
  1. DNA sample to be amplified is heated to 95ºC - break the H bonds between strands
  2. The mixture is cooled to 40-60ºC which allows primers to anneal
  3. Taq polymerase and free nucleotides added, the mixture is heated to 70ºC. Taq polymerase copies each stand and the process is repeated.
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3
Q

What are the functions of primers

A
  1. Stop the 2 DNA strands rejoining
  2. Bracket the section of DNA to be copied
  3. DNA replication can only start with a double stranded region
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4
Q

What is the role of PCR

A

Forensics, fingerprinting

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5
Q

What are DNA probes used for?

A

Identify sections of DNA that contain a specific sequence of bases. Once the specific section is identified, restriction endonuclease enzymes cut out the required section

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6
Q

What is a probe?

A

A probe is a short single strand of DNA with a specific base sequence that will hybridise to the complementary target section if present.

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7
Q

Describe the process of genetic fingerprinting

A
  1. Extracted from a DNA sample often PCR used to amplify the same
  2. The DNA is cut into fragments of different sizes by restriction enzymes. MRSs the number of repeats differs
  3. Fragments separated by gel electrophoresis
  4. Heat treated to make it single stranded
  5. Transferred to a nylon membrane and radioactive/fluorescent probes added.
  6. The labelled DNA is added to a film and is recognisable as a genetic finger print
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8
Q

What is the role of genetic finger printing in society?

A
  1. Each individual has unique MRSs which means you can identify people
  2. Establish ancestry paths
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9
Q

Describe restriction endonucleases

A
  1. They occur naturally as defence bacterial enzymes that cut up foreign DNA injected by bacteriophage
  2. Cuts the DNA via a hydrolysis reaction at a specific section of bases called a hydrolysis site
  3. Can either create blunt or sticky ends
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10
Q

What are sticky ends?

A

Short sections of DNA where there is only one strand and the bases in that section are unpaired and exposed.

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11
Q

What is the role of reverse transcriptase?

A

It is used to make the desired section of DNA from the genes mRNA

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12
Q

How does reverse transcriptase work?

A
  1. Isolating and extracting the mRNA of the desired gene
  2. Reverse transcriptase can be used to make a single strand of DNA using mRNA as a template.
  3. The enzyme DNA polymerase is used to make double stranded DNA from cDNA
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13
Q

How can differences in the nucleotide sequence be identified?

A

Gene sequencing and genetic markers

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14
Q

What are two types of genetic markers?

A
  1. MRSs - Microsatellite Repeat Sequences, junk DNA has short tandem repeats, the number of repeats is unique to each individual
  2. SNPs - Single Nucleotide Polymorphisms, A change in one nucleotide in a section of DNA can give rise to different alleles.
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15
Q

Describe plasmids as vectors

A

The bacterial plasmid is a closed loop of DNA. If cut with the same restriction endonuclease enzyme used to cut the donor DNA complementary stick ends will be made. They will match up perfectly. DNA ligase will join the two strands together due to the formation of phospho-diester bonds. The donor DNA is spliced into the plasmid and it becomes a closed loop again. The plasmid is now a recombinant plasmid.

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16
Q

Describe viruses as vectors

A

Viruses are naturally adapted to shoot their genetic material into a host cell. A bacteriophage can have donor DNA spliced into its own which is then fired into the host cell.

17
Q

How do you insert the gene into the host cell via plasmids

A
  1. Bacterial cells incubated with Calcium ions and subjected to heat shock 0ºC to 40ºC - makes bacteria more permeable
  2. Identify cells that have taken up the desired Gene - use of marker genes e.g. (1) antibiotic resistance, cuts one gene for antibiotic resistance using replica plating, Replica plating blotting the original plate, the pressing onto a new plate. (2) Using gene probes - identify if host DNA is recombinant
18
Q

What are some genetically engineered microorganisms

A
  1. Insulin - for diabetes, bacteria. Previously had to be extracted from dead animals
  2. HGH, Enzymes, Adhesives, lung surfactant protein and interferon
19
Q

How are virus’ used to target cancer cells?

A

Genetically modified so that they do not harm normal cells, provides a stronger immune response

20
Q

How do viruses treat bacterial infections?

A

When bacteria becomes resistant to antibiotics, GM viruses can support antibiotic action by disrupting these evolving defensive strategies in antibiotic resistant bacteria

21
Q

How do transgenic plants come about?

A
  1. Agrobacterium enter the plant at a point where it was damaged
  2. Gene guns - coat microscopic pellets with donor DNA which is then fired into the host cell. Hit or miss
22
Q

What are benefits of GM crops?

A
  1. Herbicide resistance
  2. Pesticide resistance
  3. Disease resistant
  4. Greater ecological range - drought tolerant
  5. Nutritional enhancement
23
Q

What are downfalls of GM crops?

A
  1. Not natural
  2. Creation of super weeds
  3. Outcompete non enhanced species
  4. Can cause allergies
24
Q

How is donor DNA incorporated into animals?

A
  1. Use of liposomes - use of a lipid carrier allows it to cross the lipid bilayer
  2. Electroporation - Disrupt cell membrane with high voltage
  3. Viruses
  4. Insert host DNA directly into the fertilised eggs of an animal
25
Q

What are some advantages of GM animals?

A
  1. Encourage faster growth and better quality

2. Produce medicinal substances

26
Q

What is the principal of gene therapy?

A

The functional allele can be inserted into the cells affected by the condition

27
Q

What are the two types of gene therapy and describe

A
  1. Somatic-cell - Only targets the affected tissues. only affects specific and easily reached parts of the body e.g. lungs. Can be used at any stage in an individuals life
  2. Germ-line - Replacing the defective genes in the fertilised egg. The defective gene will not pass to their offspring as it is fixed.
28
Q

Which is used

A

Only somatic cell is used currently as the technology is not available for germ line and it is immoral as fuck

29
Q

How is the functional DNA transferred into the recipients cells during somatic cell gene therapy

A
  1. Adenoviruses - they are able to inject their DNA into lung epithelial cells. Harmful DNA needs taken out and donor DNA needs spliced in.
  2. Retroviruses - The RNA and donor RNA is converted by reverse transcriptase into DNA in the host cell
  3. Liposomes - artificial lipid vesicle encapsulates the donor DNA, goes into host DNA
30
Q

What is the genome?

A

The complete nucleotide base sequence of an organism

31
Q

What are genome sequencing and genetic mapping?

A

Genome sequencing - Working out the base sequence of an organism
Genetic mapping - Identifying where particular genes are on chromosomes

32
Q

Genome sequencing - key steps

A

Yeast was the first eukaryotic cell to be fully sequenced
Round worm was the first multicellular organism to be sequenced
Over time it became faster and cheaper

33
Q

What is the human genome project?

A

Set up to map and sequence 3 billion nucleotides in the human genome and to identify all genes present
Just over 21,000 genes

34
Q

What is microarray technology?

A
  1. Enables the analysis of genes
  2. Contains a base of many wells each with a different sequence of DNA
  3. Single stranded DNA is washed over the array. Any complimentary DNA will hybridise and produce a fluorescent tag.
  4. The strength of the tag indicates the level of gene expression
35
Q

What is microarray technology used for?

A

Identify mutations, SNPs or provide information of the expression levels of genes

36
Q

What are the benefits of genome sequencing?

A
  1. More detailed mapping of genes, identify harmful carriers
  2. Drug development - pharmacogenetics - designer drugs - personalised medicine. medicine that is specific to the genome of individuals
  3. Increased potential of gene therapy
  4. Enables the primary structure of proteins to be worked out
  5. Find out ancestral links and evolutionary development
37
Q

What is a gene knockout?

A

A transgenic organism in which a a gene has been removed

38
Q

What is a gene knock in

A

Where a gene has been added in, study disease progression

Biology Module 2 (5 decks)

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