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Biology > Mr Dewhurst- manipulating genomes > Flashcards

Mr Dewhurst- manipulating genomes Flashcards

1
Q

what is a genome

A

all genetic information of an individual, population or species

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2
Q

what is genomics

A

study of genomes

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3
Q

what is gene technology

A

manipulation of DNA

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4
Q

what are autosomes

A

chromosomes that aren’t sex related

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5
Q

what is DNA profiling

A

producing an image of patterns in non-coding DNA in individuals

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6
Q

what is the method of DNA profiling known as

A

Sanger method

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7
Q

what is the first stage of DNA profiling

A

extract DNA from tissue sample and then using a technique called PCR can produce enough DNA to profile

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8
Q

what is the second stage of DNA profiling

A

digesting sample- strands of DNA are cut into smaller fragments using enzymes called restriction endonucleases - each enzyme makes one cut through each strand of DNA on helix
different enzymes cut a specific nucleotide sequence

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9
Q

what is in the test tube with DNA fragments

A

primers, terminator bases ( stop DNA synthesis)

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10
Q

what is the third stage of DNA profiling

A

separating DNA fragments using electrophoresis which uses an electric current to separate fragments as phosphate on fragment ( nucleotide) is negatively charged so attracted to cathode at bottom.

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11
Q

what is the fourth stage of DNA profiling

A

hybridisation - fluorescent DNA probes or radioactive labels added to join to complementary sequence so fragments are visible

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12
Q

what is the last stage of DNA profiling

A

seeing evidence- if radioactive labels added first then x-ray images produced, if fluorescent dies added then it is viewed under UV rays
the shorter fragments will be at the bottom

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13
Q

what is the purpose of Polymerase chain reaction

A

to amplify millions of copies of DNA - can be used for paternal testing, crime scenes, identifying new species

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14
Q

what is the first stage of PCR

A

denature DNA by heating machine up to 95* which breaks hydrogen bonds between DNA strands so they separate

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15
Q

what is the second stage of PCR

A

annealing of primers- machine cooled to 55* to allow primers to join which prevents strands joining back together and signals DNA polymerase where to begin replication

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16
Q

what is the third stage of PCR

A

synthesis of DNA- temperature increased to 72* to allow taq polymerase to catalyse the formation of sugar phosphate backbone

17
Q

why is PCR good for crime scenes

A

often little traces of DNA found so not enough to provide evidence for analysis but can be copies and amplified for analysis

18
Q

what are the 2 principles of genetic engineering

A

genetically modifying DNA by combining DNA from different organisms to produce recombinant DNA
genes are isolated from one organism and inserted into another organism using a vector

19
Q

what is the summarised process of genetic engineering

A

1- gene isolated
2- copy of gene placed in vector
3- vector carries gene into recipient cell
4- recipient expresses novel gene ( new gene)

20
Q

what is one way of isolating genes

A

extract mRNA from cells to show what gene is expressed, reverse transcriptase then copies mRNA back to DNA called cDNA (complementary DNA) so it can be copied into double stranded DNA which codes for original protein

21
Q

what is another way of isolating genes

A

restriction endonuclease - cut gene staggered leaving some bases unpaired called sticky ends

22
Q

what is it called when sticky ends join together

A

ligation

23
Q

outline the process of inserting gene into a cell using a plasmid

A

1- chromosomal DNA of organism A is extracted
2- gene of interest is amplified using PCR A
3- multiple copies of gene is made from organism A
4- gene is inserted into bacterial plasmid using ligase enzyme
5- plasmid now contains gene from organism A
6- the plasmid is then inserted into organism B

24
Q

what is one way of getting the vector (plasmid) into recipient cell

A

heat shock- heats host cell from 0-42* in the presence of calcium ions to allow the membrane to become more porous
the CA2+ surrounds DNA to allow it to enter through phospholipid bilayer and foreign DNA enters cell

25
Q

what is a problem with heat shock

A

not all bacteria plasmids take gene so not all recipient cells have the required gene

26
Q

what is a way of solving the problem with heat shock

A

add antibiotic resistance as well as gene, so the ones that grow have the gene needed as the ones without the gene will respond to antibiotics and not grow

27
Q

what is transfection

A

electrofusion- electrical fields help to introduce DNA into cells
T1 (recombinant) plasmids - inserted into bacterium called agrobacterium tumefaciens, which infects some plants and naturally inserts its genome into host cell

28
Q

what is somatic cell gene therapy

A

replacing mutated body cells with healthy body cells.

29
Q

what are the uses of both types of gene therapy

A

stop disease in embryos, cures conditions/diseases

30
Q

what are 3 methods of getting genes into cells

A

vector( viral vectors), plasmids , lysosomes

31
Q

why has gene therapy had limited success so far

A

hard to control where it is inserted so could disrupt other cells causing them to divide uncontrollably- cancer
somatic- has to be regularly redone to millions of cells as body cells regenerate
hard to get alleles into nucleus without the immune system fighting them as being foreign cells.

32
Q

what is germline cell therapy and some drawbacks

A

replacement of embryonic mutated cells with healthy ones- last whole lifespan
illegal, could be used wrongly- design a baby, embryo cannot consent

33
Q

what are the two types of somatic gene therapy

A

In vivo – the new gene is inserted via a vector into cells inside the body
Ex vivo – the new gene is inserted via a virus vector into the cell outside the body. These cells are then grown in the laboratory and returned to the person by an injection into a vein

34
Q

what are some reasons for genetic engineering of plants and animals

A

increases yield- more food supply for growing population
pharmaceutical substances- treat diseases
reduce use of pesticides- increases biodiversity
grown in harsh conditions

35
Q

what are some reasons against genetic engineering

A

private companies will increase the prices that small business cannot afford due to large machinery needed- they get less business
further health problems such as allergies- not known long term effects
animal cruelty- goes against personal and religious beliefs

Biology (18 decks)

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