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Biology > Topic 8 > Flashcards

Topic 8 Flashcards

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1
Q

Isolating target gene stages

A

1) restriction enzymes
2) reverse transcription
3) artificial synthesis gene

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2
Q

Isolating target genes - restriction enzymes

A
  • DNA contains palindromic sites
  • restriction enzymes cut DNA at specific palindromic sites called restriction sites
  • if theres a restriction site either side of target gene
  • restriction enzymes can be use to cut it out
  • using restriction enzymes leaves DNA with sticky ends
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3
Q

Isolating target genes - reverse transcriptase

A
  • cells only have 2 copies of each gene (in the nucleus), it is hard to access
  • enzyme that does transcription backwards
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4
Q

Isolating target genes - artificial synthesise gene

A
  • use a ‘gene machine’ to make DNA from scratch
  • join about 25 nucleotides together at once
  • forms an oligonucleotide
  • join oligonucleotides together to form a synthetic gene
  • design your own gene
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5
Q

Inserting target gene stages

A

1) isolating target gene
2) insert gene into a vector
3) insert vector into bacteria

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6
Q

Inserting target genes - isolating target gene

A
  • through a: gee machine, reverse transcriptase, restriction enzymes
  • needed to add: a promoter region, terminator region, sticky ends, marker gene
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7
Q

Inserting target genes - insert gene into a vector

A
  • use the same restriction enzymes to cut plasmid (sticky ends are complementary
  • DNA ligase reform the phosphodiester bonds
  • forms recombinant DNA
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8
Q

Vector

A

Something that’s used to move DNA from one place to another

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9
Q

Recombinant DNA

A

DNA from more than one source/ organism

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10
Q

Inserting target genes - insert vector into bacteria

A
  • transgenic organism - contains recombinant DNA
  • ice cold calcium chloride
  • heat shock (increases permeability of bacterial cell wall)
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11
Q

Marker genes

A

Genes that are paired with target genes to check if the vector has been inserted properly

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12
Q

Process of marker genes

A
  • vector are often not take up by bacteria
  • to tell which bacteria transformed you need marker gene
  • transformed bacteria contains the recombinant DNA (target + marker gene)
  • Marker genes can be easily identified
  • only bacteria that have accepted the vector (transgenic bacteria) are selected and cultured
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13
Q

UV fluorescence as marker genes

A

Will fluoresce under UV

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14
Q

Antibiotic resistance as a marker gene

A

Will be able to survive in a culture with antibiotic

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15
Q

What is the use of PCR

A
  • used to amplify DNA

- sometimes called in vitro DNA amplification

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16
Q

What is needed for PCR

A
  • DNA sample
  • free DNA nucleotides
  • primers (need to select which part of DNA is copied)
  • DNA polymerase
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17
Q

What are primers

A

Short sequences of DNA that are complementary to the start of DNA sample

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18
Q

Steps of PCR

A

1) heat to 95C
2) cool to 50C
3) heat to 70C (allows rate of reaction to happen fast)
4) repeat

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19
Q

Why do you heat up to 95C for PCR

A
  • break H bonds

- Make DNA single stranded

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20
Q

Why do you cool sample to 50C in PCR

A
  • allows primers to bind
  • complementary base pairing
  • DNA double stranded
  • DNA polymerase can bind
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21
Q

Why do you heat sample to 70C in PCR

A
  • DNA polymerase adds complementary nucleotides

- forms phosphodiester bonds

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22
Q

Why do you repeat the PCR method

A

each cycle we double the DNA

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23
Q

Summary of gene technology stages

A

1) isolate target gene
2) insert gene into vector
3) insert vector into bacteria
4) identify transgenic organism
5) culture transgenic bacteria
6) extract + purify protein

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24
Q

What do you use to isolate target gene

A
  • restriction enzymes

- (gene machine, reverse transcription, promoter + terminator)

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25
Q

What is used to insert a gee into a vector

A
  • same restriction enzyme
  • sticky ends to be complementary
  • DNA ligase
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26
Q

What is used to insert vector into a bacteria

A
  • recombinant DNA

- Ice cold CaCl2 + Heatshock

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27
Q

What is used to identify transgenic organisms

A
  • marker gene

- U.V. Fluorescence/ radioactivity/ antibiotic resistance

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28
Q

What is used to culture transgenic bacteria

A
  • transcribe + translate recombinant DNA

- Make the protein of target gene

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29
Q

Gene therapy

A

Changing faulty alleles that cause genetic disease

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30
Q

Gene therapy for dominant alleles

A
  • sufferer will be heterozygous
  • they will have the functional allele
  • silence dominant allele
  • use a vector to add a DNA fragment into dominant allele
  • dominant allele won’t be transcribed
  • recessive allele expressed
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31
Q

Gene therapy for recessive alleles

A
  • sufferer will be homozygous
  • use a vector to add the functional allele to DNA
  • dominant allele will be expressed
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32
Q

Possible problems with gene therapy

A
  • allele insert into wrong locus
  • could silence wrong gene by mistake
  • Gene could be over expressed
  • use of gene therapy could be used for non-medical uses
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33
Q

Germ line gene therapy

A

Change the allele of gametes

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34
Q

Inheritance from germ line gene therapy

A
  • all future offspring inherit

- illegal in humans

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35
Q

Somatic gene therapy

A

Changing the alleles of body cells (non gametes)

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36
Q

Inheritance from somatic gene therapy

A

Offspring do not inherit change

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37
Q

Mutation

A

A change to the nucleotide sequence of DNA

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38
Q

Mutagenic agent

A

Increase the rate of mutations

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39
Q

Base analogs

A

A chemical that can substitute from a normal nucleotide base

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40
Q

Radiation

A

Change the structure of DNA

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41
Q

Change DNA bases

A

Chemicals that react with DNA to change its structure

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42
Q

Substitution

A

One base is swapped for another

- no change, single amino acid changed

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43
Q

Addition

A

An extra base is added

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44
Q

Deletion

A

A base is removed

- frame shift, all the following triplets in the sequence will be affected

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45
Q

Inversion

A

A sequence of bases is reverse

- no change, change a few amino acids, no frame shift

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46
Q

Duplication

A

One or more bases are repeated

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47
Q

Translocation

A

A sequence of DNA is removed from one part of the genome and moved to another part of the genome

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48
Q

Changes to the base sequence of DNA

A
  • no change to amino acid sequence
  • change to a single amino acid
  • change to sequence of many amino acids
49
Q

Why is there no change to amino acid sequence after a mutation

A
  • amino acids are coded for by more than one triplet on DNA

- it is degenerate

50
Q

Why is there only change to a single amino acid after a mutation

A
  • changes to one DNA triplet
  • changes the translation of one amino acid
  • changes primary structure of the protein
  • change the hydrogen/ ionic bonding
  • change the tertiary structure of the protein
51
Q

Why is there a change to the sequence of many amino acids

A
  • frame shift changes the sequence of all the following DNA triplets
  • change all the following amino acids in sequence
  • changes the primary structure
  • changes the hydrogen/ ionic bonding
  • change tertiary structure
52
Q

Stem cells

A

Can divide and differentiate to become different types of cell

53
Q

Totipotet

A

Stem cells that can differentiate into any type of specialised cell

54
Q

Where are totipotent cells from

A
  • early mammalian embryos

- can form placental cells

55
Q

Pluripotent

A

Stem cells that can differentiate into many types of specialised

56
Q

Where are pluripotent stem cells from

A
  • embryos

- can’t make placental cells

57
Q

Multipotent

A

Stem cells that can differentiate into a few types of specialised cells

58
Q

Example of multipoint cells

A

bone marrow to RBC and WBC

59
Q

Unipotent

A

Stem cells that can differentiate into one type of specialised cell

60
Q

Example of unipotent cells

A

Heart unipotent cells can make cardiomycetes only

61
Q

Cell specialisation/ differentiation

A
  • all cells contain 100% of an organisms DNA
  • conditions within cells control which genes are expressed into proteins
  • change internal environment of the cell - affect the expression of other genes (are specialised)
62
Q

iPs cells

A

Treating unipotent stem cells with transcription factors that make them become pluripotent

63
Q

How to make induced pluripotent stem cells

A
  • use a modified virus as a vector
  • virus inserts transcription factor genes from pluripotent cells into the DNA of unipotent stem cells
  • transcription factors are expressed
  • ultimate goal is to make totipotent stem cells this way
64
Q

Creation of embryonic stem cells

A
  • embryos are made in a lab by IVF
  • pluripotent stem cells are removed after a few days
  • embryo is destroyed
  • pluripotent stem cells can differentiate into all types of body cells
65
Q

Creation of adult stem cells

A
  • taken from adult in an operation
  • adult stem cells are multipoptent
  • less useful for medicine as they can’t form every type of cells
66
Q

Why are stem cells used

A

They can differentiate to form any type of specialised cell

67
Q

Bone marrow transplants

A
  • contains multipotent stem cells
  • can differentiate into RBCs and WBCs
  • if someone has fault bone marrow you can replace there bone marrow to form new healthy cells
68
Q

Growing new organs with stem cells

A
  • no donors needed
  • iPs cells
  • iPs organs have the same antibodies so there is no organ rejection
69
Q

Advantages of stem cells in medicine

A
  • saves lives
  • improve quality of life
  • prevent suffering
70
Q

Ethical issues with stem cells in medicine

A
  • they are taken from IVF embryos which could be developed into a foetus if implanted
  • some people believe from fertilisation a zygote has a right to life
  • adult stem cells are not pluripotent
  • iPs cells could be the future solution
71
Q

Transcription factors

A

Proteins that control the rate of protein synthesis by switching some genes on and other genes off

72
Q

Promoter region

A

Short sequence of DNA at the start of a gene

73
Q

Stages of activators and repressors

A

1) transcription factor moves from the cytoplasm into the nucleus
2) binds to the promoter region
3) activators - help RNA polymerase to bind to DNA ( gene is transcribed)
4) repressors - prevent RNA polymerase binding to DNA (gene is not transcribed)

74
Q

Controlling transcription factors - oestrogen

A
  • oestrogen is a steroid hormone
  • oestrogen binds to a receptor called an oestrogen receptor
  • forms oestrogen-oestrogen receptor complex
  • transcription factors can also be turned on/off by second messengers
75
Q

Cancer

A

Uncontrolled cell division

76
Q

How do tumour suppressor genes work

A
  • proteins that slow down the rate of mitosis or speed up the rate of apoptosis
  • if theres a mutation into the tumour suppressor gene
  • the protein might be non-functional
77
Q

How to proto-oncogenes work

A
  • make proteins that increase the rate of mitosis
  • if a proto-oncogene mutates it is called an oncogene
  • oncogenes can be over expressed
78
Q

Epigenetics

A

Changes to gene expression caused by environmental factors

79
Q

How does epigenetics work

A
  • controls gene expression by preventing transcription
  • can be inherited between generations
  • helps organisms respond to changes in their environment
80
Q

Methylation of DNA

A

More Methylation = Terminate Transcription

  • methyl can attach to DNA at CPG sites
  • Methylate CpG sites prevent transcription enzymes attaching
  • transcription is prevented
81
Q

Affects of acetylation of histones

A

Less acetylation of histones Terminates Transcription

82
Q

How does the acetylation of histones work

A
  • acetyl groups make histones space out
  • allows transcriptional enzymes to attach - gene is expressed
  • enzymes remove acetyl groups - prevents transcription
83
Q

What does RNAi do

A

Controls gene expression by preventing translation

84
Q

siRNA - small interfering RNA

A
  • short double stranded RA
  • combines with proteins to form a siRNA-protein complex
  • siRNA is single stranded
  • siRNA has complementary base sequence to target mRNA
  • siRNA protein complex breaks down the mRNA into pieces
  • prevents translation
  • mRNA pieces are recycled
85
Q

MicroRNA

A
  • microRNA combines with a protein to form a microRNA-protein complex
  • bind to mRNA by complementary base pairing
  • less specific that siRNA - works on more than one mRNA
  • prevents translation by stopping the ribosome attatching
  • mRNA can be stored and used later or recycled
86
Q

Hypermethylation

A

Tumour suppressors genes

- means that proteins not transcribed

87
Q

Hypomethylation

A

Proto-oncogenes

- more protein is transcribed

88
Q

Benign tumours

A
  • non-cancerous
  • grow slowly
  • harmless
  • can become malignant
89
Q

Malignant tumours

A
  • are cancerous
  • grow quickly
  • destroy tissue
  • can break up and spread in blood/ lymphatic system
90
Q

Oestrogen and breast cancer

A
  • high levels of oestrogen can cause some types of breast cancer
  • oestrogen can bind to proteins to form a transcription factor called oestrogen-oestrogen receptor complex
  • increase rate of cell division, increase DNA replication, increase mutation, increase cancer
91
Q

Identifying tumour cells

A
  • mitosis - more cells dividing
  • nuclei - large and odd shaped, more than 1 per cell
  • irregular cell shape
  • loss of normal function
  • disorganised arrangement
92
Q

Genome

A

The complete set of genetic material that an organism has

93
Q

Proteome

A

The complete set of protein that an organism can make

94
Q

Sequencing projects

A
  • human genome project - sequenced whole human genome
  • only sequence short fragments at once
  • split genome up into small sections -> sequence -> put back together
  • sequencing methods are constantly being up dated
95
Q

Predicted amino acid sequence - simple organisms

A
  • few regulatory genes

- little non-coding DNA

96
Q

Predicting amino acid sequence - complex organisms

A
  • regulatory gene turns on/off other genes
  • lots of non-coding DNA
  • hard to determine proteins from DNA
97
Q

Uses of genome projects

A
  • understanding evolutionary relatedness

- medicine - understand the antigens to develop new vaccine

98
Q

uses of genetically modified organisms in agriculture

A
  • express protein from bacteria
  • protein is toxic to insects
  • fewer insects eat soya plants
  • gene from corn to rice expresses vitamin A
99
Q

advantages of using genetically modified organisms in argiculture

A
  • uses less chemical pesticides
  • more efficient food chain
  • prevents blindness caused by vitamin A deficiency (corn to rice gene)
100
Q

disadvantages of using genetically modified organisms in agriculture

A
  • monocultures have a low genetic diversity
  • susceptible to environmental factors eg disease
  • have to buy seeds every year
  • decrease in biodiversity
101
Q

uses of genetically modified organisms in industry and research

A
  • making enzymes eg lipase

- transformed pathogen to TREAT disease - attach other pathogens but don’t infect humans

102
Q

advantages of using genetically modified organisms in industry and research

A
  • reduce energy and cost
  • fast and cheap production
  • treat disease
  • pathogens won’t develop resistance
  • reduce suffering
103
Q

disadvantages of using genetically modified organisms in industry and research

A
  • could mutate and infect humans

- could be used in a negative way

104
Q

uses of genetically modified organisms in medicine

A
  • transform bacteria to express proteins

- mammals can also be transformed to produce useful products in their milk

105
Q

advantaged of genetically modified organisms in medicine

A
  • makes human proteins

- cheaper and easier than making protein synthetically

106
Q

disadvantages of using genetically modified organisms in medicine

A
  • possible unexpected problems

- using animals as commodities

107
Q

DNA probes

A

a short sequence of DNA that is complementary to a specific allele/ mutation / gene

108
Q

uses of DNA probes

A

to test if a sample of DNA contains a specific sequence

109
Q

stages of using a DNA probe

A

1) attach a label to DNA probe
2) is sequence is present than DNA probe will hybridise and stick to DNA sample with a complementary sequence
3) rise to remove unhybridized DNA probes
- DNA must be single stranded

110
Q

DNA microarray stages

A

1) many DNA probes attached to a tile in a grid
2) add patients DNA with label
3) DNA will hybridise to complementary DNA
- 4) rinse and view labels

111
Q

How do you use DNA microarry

A
  • attach the label to human DNA

- test multiple alleles at once

112
Q

DNA probes in genetic counselling

A
  • used to identify: carriers of a genetic disease, specific alleles, the most effective treatment
  • healthcare professionals can advise people about the risks of passing on inheritable diseases, developing diseases later on in life, making informed decisions about treatment and preventions
113
Q

uses of DNA probes in genetic screening

A
  • parents can see if they are carriers of recessive alleles

- you can diagnose and treat before symptoms show

114
Q

uses of DNA probes in personalised medicine

A
  • if a doctor knows a patients genotype it gives the best drugs for them
  • change drugs if you know a patient has an allele that causes a side effect with the normal drug
115
Q

genetic fingerprinting

A

identifying individuals by comparing the differences in their Variable Number Tandem Repeats (VNTR)

116
Q

genetic fingerprinting in forensics

A
  • sample of DNA from a crime scene and suspects
  • amplify using PCR
  • run gel electrophoresis
  • the chances of two individuals having the same number of VNTR’s is very low
  • suspects will have more matches to the crime scene
117
Q

genetic fingerprinting for relatedness

A
  • the more closely related two individuals are the higher the percentage match of VNTRs
  • conservation of endangered species to avoid inbreeding by mating individuals who are least related to help maintain genetic diversity
118
Q

genetic fingerprinting for medical diagnosis

A
  • genetic fingerprints can be used to test for specific combinations of alleles
  • can be used to diagnose genetic disorders
119
Q

facts of genetic fingerprinting

A
  • non-coding DNA contains lost of VNTRs
  • don’t code for proteins
  • don’t affect phenotype - vary more than coding DNA
  • the number of repeats of each VNTR varies

Biology (18 decks)

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