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O'Donovan Alevel bio > Ch21 > Flashcards

Ch21 Flashcards

1
Q

conversion of mRNA to cDNA

A

reverse transcriptase cause MRNA to act as template for complementary nucleotides to bond and form cDNA

cDNA hydrolysed and becomes single-stranded

DNA polyermase builds up complementary nucleotides to make double-stranded

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2
Q

Isolating gene

A

Use restriction endonuclease enzyme
Cuts DNA at specific base sequence(recognition sites)
Creates sticky ends
Sticky ends are palindromic and so are complementary base sequences

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3
Q

Gene machine advantage

A

Faster as enzyme-catalysed reactions have extra steps

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4
Q

Gene machine steps

A
  1. identify amino acid sequence of desired protein
  2. work out mRNA and DNA sequence
  3. enter DNA sequenece into computer which checks biosafety
  4. computer creates small sections of overlapping single strands of nucleotides(oligonucleotides)
  5. each oligonucleotides join together to make gene
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5
Q

Advantage of reverse transcription

A

cDNA is intron free because it comes from mRNA template

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6
Q

vector

A

transfer genes from one organism into bacteria host cell

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7
Q

Overall process of making protein

A
  1. isolation of DNA fragments that have desired protein gene
  2. insertion of DNA into vector
  3. transformation of DNA into host cells
  4. identification of host cells using gene marker
  5. growth/ cloning
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8
Q

in vivo DNA insertion

A

Restriction endonuclease cuts plasmid
Restriction endonuclease cuts and isolates desired gene
Same enzyme cuts at same base sequence
Plasmid and gene DNA fragments have complementary sticky ends that bond
DNA ligase anneals gene and plasmid together through forming phosphodiester bonds

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9
Q

Before insertion of DNA frament

A

add promoter and terminator region so synthesis of mRNA can occur
RNA polymerase knows when to begin and end transcription

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10
Q

in vivo cloning

A
  1. add marker gene(antibiotic resistance or fluorescent)
  2. mix plasmid and bacterial cells in medium with calcium ions so plasmids can insert themselves
  3. put bacteria colony onto medium where marker gene is expressed
  4. bacteria that remain resistant or become fluorescent contain desired gene
  5. purify and clone
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11
Q

Marker genes

A
  1. antibiotic resistant
  2. makes fluorescent protein
  3. produces enzyme with particular action
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12
Q

Polymerase chain reaction

A
  1. DNA heated to 95
  2. strands separate
  3. cooled to 55
  4. primers anneal to complementary base pairs
  5. nucleotides attach by complementary base pairing
  6. temp raised to 75
  7. DNA polymerase(binds to primers) and bonds nucleotides together
  8. repeat the cycle
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13
Q

Primer

A

short sequences of nucleotides that have set bases complementary to those at one end of each of 2 DNA fragments

starting place for DNA polymerase
prevents 2 strands from re-joining

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14
Q

in vitro gene cloning advantage

A
  1. very rapid
  2. doesn’t require living cells
  3. creates large amount of DNA
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15
Q

in vivo cloning advantage

A
  1. able to introduce gene into another organism using vector
  2. no risk of contamination
  3. accurate
  4. cuts out specific genes
  5. transformed bacteria can produce large amount of desired protein
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16
Q

DNA probe

A

has complementary base sequences to desired allele base sequence

allows allele to be identified- fluorescent after binding or radioactive under XRAY

17
Q

DNA hydrbidisation

A
  1. heat DNA until it separates into single strands
  2. cool
  3. DNA probe anneals to complementary bases sequence of allele
  4. DNA cleaned
  5. hybridised DNA is now labelled and can be identified
18
Q

Genetic screening

A
  • locates mutant alleles
  • locate oncogenes
    -locate mutant tumor supressor genes
19
Q

Personalised medicine

A

individuals genotype can determine effectiveness and dosage of particular drugs

20
Q

Genetic counselling

A

Family history of inherited disease and likelihood of inheritance

21
Q

Genetic fingerprinting

A
  1. extract DNA from sample and increase amount by PCR
  2. Cut DNA into smaller fragments using restriction endonuclease
  3. Separate DNA fragments using gel electrophoresis
  4. Put sample into wells on gel sheet, pass electric current through
  5. immerse gel in alkaline solution
22
Q

Gel electrophoresis

A

Place DNA fragments sample into wells of a gel plate
Apply electrical current
Immerse gel in alkaline solution
DNA fragments will travel towards positive charge
Smaller fragments will travel further

23
Q

Uses of genetic fingerprinting

A
  1. medical diagnosis
  2. breeding
  3. genetic relationships
  4. forensic science
24
Q

Variable number tandem repeats

A

repetitive non-coding bases of DNA
closer related- more identical sequences of VNTRs

O'Donovan Alevel bio (12 decks)

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