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8. Control Of Gene Expression > Genome Projects + DNA Technology > Flashcards

Genome Projects + DNA Technology Flashcards

1
Q

What is the human genome project

A

A project which successfully determined the sequence of bases of a human genome

= aim to sequence entire human Genome

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2
Q

What is the genome

A

Complete set of genes present in an organism

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3
Q

How is the human genome project used in medical advances

A

Genome projects identify which genes are responsible for certain inherited diseases = once genes are determined potential targets for drug treatment can be identified

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4
Q

What is genetic testing/screening

A

Used to identify if an individual has a specific gene that may cause a disease

Done with DNA probes complimentary to genes of diseases

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5
Q

What is gene therapy

A

Used to replace defective genes with normal,healthy genes

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6
Q

How can genome projects be used in evolutionary relationships

A

Make comparisons between individuals or species = genetic similarities can help us to show evolutionary relationships

  • can also be used in forensic testing + genetic
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7
Q

Why have we sequenced the genome of other organisms such as zebra fish

A

To identify useful genes = may help us with our own problems

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8
Q

Recombinant DNA technology

A

Transfer of DNA fragments from one organism to another

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9
Q

Why can transferred fragments in Recombinat DNA technology be translated within recipient organism

A

DNA is universal

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10
Q

Name 3 ways fragments can be produced

A

Reverse transcriptase
Restriction endonuclease
Gene machine

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11
Q

What happened after fragments are produced

A

They need to be amplified
Either by :
1- in Vitro
2- in Vivo

(Vinland saga)

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12
Q

Producing fragments - Reverse transcriptase

A

mRNA converted to DNA by enzyme Reverse transcriptase = produces cDNA (complimentary DNA)

mRNA is more accessible than DNA + there is usually a lot of it within a cell

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13
Q

Producing fragments - Restriction endonucleases

A

Restriction endonucleases cut DNA at specific base sequences known as RECOGNITION SITES = allows desired gene to be cut out of DNA and isolated

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14
Q

What 2 types of fragments can restriction endonuclease produce

A

Blunt end = cut at the same location = no exposed bases

Sticky end = staggered ends with exposed DNA bases
- the exposed ends are Palindromic and read the same forwards on one end and backwards on another

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15
Q

Producing fragments - Gene machine

A

DNA sequence entered into a computer = computer synthesis DNA strand

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16
Q

Procedure of the gene machine

A

1- If we know the desired protein amino acid sequence = work out RNA and DNA sequence
2- info fed into computer = computer checks sequence for safety to make sure we don’t produce DNA for pathogen
3- computer then forms desired gene from series of Oligonucleotides (single strands of nucleotides)

4- PCR used to amplify + make double stranded

17
Q

One advantage and disadvantage of reverse transcriptase

A

+ mRNA in cell is from the desired gene and there is lots of it

  • more steps = more time consuming
18
Q

One advantage and disadvantage of restriction endonucleases

A

+ sticky ends on DNA fragment makes it easier to insert to make recombinant DNA

  • may contain introns
19
Q

One advantage and one disadvantage of gene machine

A

+ can design exactly fragment desired with sticky ends, labels and preferred codons - do not need an organism

  • need to know the sequence of amino acids or bases
20
Q

Steps of “in vitro” amplification = PCR

A

1- DNA denatured + hydrogen bonds broken = 94• temp
2- cooled to 50• + primers added to stop DNA rejoining + to initiate base pairing of new DNA nucleotides
3- heated to 74• + taq DNA polymerase (from a thermophile bacteria) forms new strand of DNA
4- cycle repeated

21
Q

Why is taq DNA polymerase used

A

Able to withstand temps of up to 80• from a thermophile bacteria

22
Q

Steps of “in vivo” amplification = vectors

A

1- vector DNA and DNA to be inserted are cut with the same restriction endonuclease = 2 strands are complimentary
2- DNA is annealed(cooled) with DNA Ligase = strands bind together
3- Vector DNA inserted into bacterial cell = transformer cell
4- promoter + terminator regions added to DNA to be cloned

  • promoter regions stimulate RNA polymerase to transcribe this gene = producing desired protein
  • terminator regions show where the DNA polymerase can stop transcribing = end of desired gene
23
Q

What is added to genetically modified cells to show whether in vivo cloning/amplification has worked

A

Marker genes
Can code for fluorescence or antibiotic resistance

24
Q

What vectors are usually used in “in vivo” cloning/amplification

A

Bacterial plasmids
Once fragment is in vector THEN it is placed into host eg: bacterial cell
So it can be transcribed

The vector and Host are different things

25
Q

3 concerns of recombinant DNA

A

Agriculture = super weeds may evolve

Industry = may force small companies out of business through genetically modified crops + ppl may not get a say in whether they eat GM food

Medicine = designer babies

26
Q

DNA probe

A

Single stranded DNA complimentary to a specific base sequence

27
Q

What are DNA probes used for and how do they do this

A

Used to detect genes or harmful genes

Probe is complimentary to harmful gene = binds to gene in proscess known as DNA hybridisation

Probe is either fluorescently or radioactively marked

28
Q

What do genetic counsellors do

A

Give advice to people on genetics

  • give advice to parents concerned whether they will pass on faulty alleles
  • advise women identified with carrying breast cancer gene
29
Q

What are VNTRs

A

Variable number tandem repeats
Repeated base sequences that do not code for a protein

30
Q

Why can VNTRs be used in genetic fingerprinting

A

Everyone has VNTRs but the number of repeats is unique for a person

Chance that 2 people have the same number is incredibly small

31
Q

What is hybridisation and annealing

A

Hybridisation = 2 complimentary base sequences binding

Annealing = to cool /cooling such as in PCR

32
Q

Steps of genetic fingerprinting (simple)

A
  1. Extraction = extract DNA from cell + amplify via PCR due to amount of DNA being small
  2. Digestion = restriction endonucleases cut DNA into fragments
  3. Separation = fragments are separated by size with gel Electrophoresis
  4. Hybridisation = radioactive or fluorescent probes added + bind to specific Fragments
  5. Development = depends on type of probes
    Radioactive = X-ray film put over nylon membrane = radiation makes series of bars where fragments are

Fluorescent = visually can see bars

33
Q

Stages of gel electrophoresis

A
  1. DNA extracted from individual + amplified
  2. DNA is cut into fragments + Flourescently labelled
  3. Fragrant inserted into wells in gel + covered in electrically conducting buffer
  4. DNA is negatively charged = when current applied DNA moves towards positive electrode
  5. Ladder forms and current stopped after certain amount of time = smaller fragments move further
    6.exposed to UV light to show fragments
  • the lengths of DNA fragments determined by number of repeats in a VNTR = this is unique per person = DNA fragments in an individual will move different distances = ladders are unique per person

The ladder = genetic fingerprint

34
Q

Gene therapy

A

Genetic engineering technique = involves introducing new gene that translates for a desired protein that counteracts effect of disease cause by mutation

The gene introduced depends on whether mutated gene is dominant or recessive

35
Q

Is PCR a form of recombinant DNA technology

A

No = does not involve transfer of DNA between organisms

PCR is a technique used to amplify DNA sequences outside of living organisms by heating + cooling

36
Q

Somatic vs germline gene therapy

A

Somatic = alters body cells

Germline = alters sex cells (illegal)

Both involve vectors (plasmids) to insert target gene into genome of host/patient

37
Q

Following the action of reverse transcriptase is cDNA single or double stranded

A

Reverse transcriptase used the mRNA template to produce SINGLE stranded cDNA = next DNA polymerase converts single stranded to DOUBLE stranded DNA

38
Q

At what stage are promoter and terminator regions added in “vivo” cloning/amplification

A

Added to fragments of DNA before inserting them into host cells

Prompter regions have recognition sites for RNA polymerase to stimulate protein synthesis of specific protein = terminator genes are where RNA polymerase dissociates from DNA + ends transcription

8. Control Of Gene Expression (3 decks)

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