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BIOLOGY > 13.8 Genome project and gene technology > Flashcards

13.8 Genome project and gene technology Flashcards

1
Q

Genome definition

A

The complete set of genes in a cell

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2
Q

Proteome definition

A

The full range of proteins produced by cells

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3
Q

Recombinant DNA definition

A

A cell that has two or more sources of DNA

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4
Q

How is recombinant DNA achieved?

A

By isolating fragments of DNA and then inserting them into the DNA of another species

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5
Q

Universal definition

A

The same triplet coded for the same amino acid

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6
Q

Degenerate definition

A

More than one codon codes for one amino acid

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7
Q

Non overlapping definition

A

Each base is only part of one triplet

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8
Q

What are the 5 steps in recombinant DNA technology?

A
  • Isolation of genes
  • Insertion
  • Transformation
  • Identification
  • Growth/cloning
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9
Q

What are the 3 ways you can isolate a specific DNA fragment

A
  • Conversion of mRNA to complementary DNA using reverse transcriptase
  • Using restriction endonucleases to cut fragments containing the desired gene from the DNA
  • Creating the gene in a gene machine
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10
Q

Describe how to isolate a DNA fragment using reverse transcriptase

A
  • Free DNA nucleotides bind to single stranded mRNA template cia complimentary base pairing
  • Reverse transcriptase joins DNA nucleotides together to form a single stranded cDNA molecule
  • DNA polymerase is required to make cDNA double stranded
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11
Q

What are the advantages of using reverse transcriptase?

A
  • mRNA is much easier to obtain
  • Bacterial DNA doesnt contain introns
  • Using mRNA isolated from cytoplasm means introns have been removed and exons spliced back together
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12
Q

Explain how to isolate a gene using restriction endonucleases

A
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13
Q

Restrictionon endonucleases definition

A

Enzymes that hydrolyse DNA at specific recognition base sequences, usually either side of a desired gene

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14
Q

Describe isolation of DNA fragments using the gene machine

A
  • Desired nucleotide sequence fed into the computer
  • Synthesis of short sequences of nucleotides
  • Assembly of the gene using PCR
  • Gene is inserted into the plasmid
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15
Q

Vector definition

A

A DNA carrier used to transfer foreign DNA into cells

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16
Q

Describe the isolation of genes into a vector

A

Isolated target DNA fragments inserted into the vector DNA by cutting open the vector DNA using the same restriction endonucleases
This produces complimentary sticky ends between the DNA fragments ad the cut ends of the vector DNA
- Target DNA fragments anneal to the vector DNA by complimentary base pairing between the sticky ends
- DNA ligase is used to join the DNA fragment and vector at the backbone, forming phosphodiester bonds
This forms recombinant DNA

17
Q

Outline a method for in vivo cloning

A
  • Cut desired gene from DNA of desired organism
  • Use restriction endonucleases
    OR
  • Use mRNA from cell of desired organism
  • Use reverse transcriptase to form desired DNA
    THEN
  • make artificial DNA with correct sequence of bases using DNA polymerase
  • Cut plasmid open with same restriction endonucleases
  • use DNA ligase to join the sticky ends together
  • return the plasmid back to the bacterial cell
18
Q

Why do you have to identify transformed host cells?

A
  • Not all vectors take up the target DNA to become recombinant
  • Not all host cells become transformed by taking up the recombinant vectors
19
Q

Marker gene definition

A

Allows easy identification of cells that have taken up genetically transformed plasmid

20
Q

What will cells look like if they haven’t been taken up by any plasmid at all using antibiotics

A

Will be killed by both types of antibiotic

21
Q

What will cells look like that have taken up the original plasmid using antibiotics?

A

Resistant to both types of antibiotics

22
Q

What will cells look like that have taken up the transformed plasmid when using antibiotics

A

Resistant to one type of antibiotic but not the second

23
Q

How do you clone in vivo via bacteria?

A

Reproduce by binary fission

24
Q

How do you clone in vitro?

A

PCR
- polymerase chain reaction

25
Q

Polymerase chain reaction definition

A

Used to amplify DNA rapidly and efficiently

26
Q

What are the three stages of PCR

A

Seperation
Annealing
Synthesis

27
Q

Describe the process of PCR

A
  • Heat DNA to 95C to break the weak hydrogen bonds between the strands
  • Add primers and nucleotides
  • Cool to 50C to allow binding of the nucleotides
  • Add DNA Taq polymerase
  • Heat to 75C
  • Taq polymerase joins nucleotides together
  • Repeat the cycle many times
28
Q

Compare in vitro and in vivo cloning

A

In vivo compared to in vitro:
- Can be used to produce protein or mRNA from the inserted DNA as well as the target gene whereas in vitro can only be used to copy DNA
- Can be used to clone large NDA frogaments whereas in vitro gets unreliable when copying more than 1000 base pairs
- slower
Need to isolate the DNA fragment from the DNA before it can be inserted into the host cell
- Cant be used to copy partly broken down DNA

29
Q

What are the benefits of recombinant DNA technology

A
  • Develop medical applications such as insulin to treat diabetes
  • Develop agricultural applications to produce crops with a higher yield that are resistant to a disease or extreme weather
  • Better understand biological processes
30
Q

What are the concerns regarding DNA recombinant technology

A
  • Antibiotic resistance may spread
  • Inserting new genes into a plant could disrupt the other genes producing genetically modified food sources
  • Introducing herbicides could spread to wild species when they interbreed, producing super weeds
  • Improving health outcomes by recombinant DNA technology is not always available to everyone socially and economically
31
Q

Somatic gene therapy definition

A

DNA transfer to our normal body tissues

31
Q

Germ line gene therapy definition

A

DNA transfer to cells that produce eggs or sperm

32
Q

Limitations of somatic gene therapy

A
  • Not all cells take up new DNA
  • Not all cells express the DNA allele
  • Only some tissue types are accessible
    -Multiple treatments may be needed
  • Body can produce an immune response to the vector
33
Q

Gene probe

A

Short single stranded DNA molecule with a complimentary base sequence to the DNA fragment to be located which is radioactive or labelled by a fluorescent molecule

34
Q

What are the regions called that are used for DNA fingerprinting

A

Variable number tandem repeats

35
Q

Where do you find variable number tandem repeats?

A

Between genes

36
Q

Explain the process of DNA fingerprinting

A
  • Extract DNA from the nucleus of the cells
  • DNA cut into segments using restriction endonucleases
  • Must leave VNTRS intact
  • DNA fragments separated using electrophoresis
  • Mixture put into wells on gel and electric current passed through
  • Immerse gel in alkaline solution
    Radioactive marker added to hybridise the target fragments
  • Identify using autoradiography

BIOLOGY (23 decks)

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