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module 6 Genetics and ecosystems > 6.3 Manipulating genomes > Flashcards

6.3 Manipulating genomes Flashcards

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Biology 101 flashcards
1
Q

define genome

A

the complete genetic make up of an organism

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2
Q

define DNA sequencing

A

a technique allowing genes to be isolated and read

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3
Q

what did early DNA research show

A

structure of DNA was known
base triplet sequences coded for amino acids
but you couldn’t work out the sequence of nucleotide base triplets

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4
Q

explain Fred Sanger’s DNA sequencing approach

A

use a single strand of DNA as a template for 4 experiments in seperate dishes containing
one of the 4 different terminators used (modified nucleotides of ATCG)
normal nucleotides
a primer
DNA polymerase
DNA fragments of various lengths were made and pass through a gel by electrophoresis
sorted by length (shorter travelled further)
bases get read off what terminator it had and it creates the sequence

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5
Q

explain how DNA gets cloned

A

gene isolated using enzymes from bacteria
inserted into bacterial plasmid (vector) and then into an E coli bacterium host
when cultured divided lots
plasmid in DNA copied many times
each new bacteria contains DNA

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6
Q

what technique is used in high throughput sequencing to sequence genomes

A

pyrosequencing

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7
Q

what is pyrosequencing

A

synthesisng a single strand of DNA complementory to the strand to be sequenced, by one base at a time whilst detecting the light admission

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8
Q

explain the process of pyrosequencing

A

DNA cut into fragments using a nebuliser
lengths are degraded into single strands of DNA
sequencing primer is added
DNA is incubated with DNA polymerase, ATP sulfurylase, luciferase, apyrase, luciferin and APS
activated nucleotides (TTP, ATP, CTP, GTP) are added one at a time
light detected

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9
Q

what is the human genome project

A

project launched to sequence the human genome
contains about 24000 genes

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10
Q

how can genome wide comparisons be made between species

A

whole genome sequencing determins the complete DNA sequence of an organisms genome

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11
Q

how can genome wide comparisons be made between individuals

A

all have same genes but different alleles
mutations can cause difference between DNA sequences

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12
Q

what is bioinformatics

A

developing and using computer software that can analyse / store biological data

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13
Q

how can we predict the amino acid sequence of proteins

A

can work out the base triplets which code for an amino acid = can determine the orimary structure of an amino acid

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14
Q

define intron

A

non coding bit of DNA that doesn’t code for an amino acid

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15
Q

define gene

A

sectionof DNA that codes for a protein and results in a characteristic

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16
Q

define locus

A

position of a gene on a chromosome

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17
Q

what is synthetic biology

A

involves designign and building biological dveices and systems

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18
Q

uses of synthetic biology

A

information storage onto single strands of DNA
productionof medicines
development of porotiens - eg. haemoglobin
biosensors
nanotechnology

19
Q

what is DNA profiling

A

technique used to determin patterns in the non coding DNA of an individual

20
Q

explain the process of DNA profiling

A

DNA is obtained from the individual
digested with enzymes into fragments
fragments are sperated by gel electrophoresis
a banding pattern is created and can be compared

21
Q

who developed DNA profiling

A

Alac Jeffreys

22
Q

uses of DNA profiling

A

forensic science
maternity and paternity disputes
analysis of disease

23
Q

define polymerase chain reaction

A

technology that can amplify a short length of DNA to lots of copies

24
Q

process of PCR

A

DNA mixed with DNA nucleotides, primers, magnesium ions and enzyme Taq DNA polymerase
heated to 95 degrees c - breaks hydrogen bonds between complementary nucleotide base pairs into two single strands
cooled to 68 degrees c - primers can bind to one end of each single strand (gives a small section of a double strand)
DNA polymerase enzymes bind to the end where the primer is
heated to 72 degrees c - optimum temp for DNA poly
DNA poly catalyses nucleotides joining the single strand in the 5’ to 3’ direction
new double strand generated
repeats cycles

25
Q

uses of PCR

A

forensics - small quantities of DNA can be amplified
analysis of disease risk - detecting mutations
identify viral infections
tissue typing - reduces risk of rejection of transplants
detection of cancer
research of extinct species

26
Q

define electrophoresis

A

process used to seperate proteins/ DNA fragments of different sizes

27
Q

explain how electrophoresis works

A

extraction of DNA from sample
fragments are created using restriction endonucleas enzymes)
seperation of fragments by running electric current over it (attracted to positive electrode at different rates)
in an agarose gel and buffer solution
smaller fragments = travel further
fluorescent dye/ probes used to show

28
Q

how can electrophoresis be used for seperating proteins

A

charged solution to make all proteins have same charge
proteins seperate as they move through the gel
according to molecular mass

29
Q

what is a DNA probe

A

short single stranded length of DNA complementory to section of DNA being investigated
has fluorescent dye/ radioactive tag

30
Q

explain how a DNA probe is used

A

DNA probe binds to the
complemenotry section of the target DNA
the fuorescent dye tag is shown up when UV light passes over/ radioactive tag shows up dark when exposed to photographic film
identifies presence/ location of the traget DNA sequence

31
Q

uses of DNA probes

A

locate specific gene
identify same gene in variety of genomes
idenitfy presence of an allele

32
Q

what is genetic engineering

A

recombination DNA technology
combining DNA from different organisms by isolating them and inserting them into another organism using vectors

33
Q

name the stages of DNA engineering

A

obtaining required gene
placing gene into a vector
get vector into recipient cell
recipient expresses gene

34
Q

how is the required gene obtained in genetic engineering

A

mRNA from cells where gene is expressed is obtained
enzyme reverse transcriptase catalyses formation of single strand of complementary DNA
primers and DNA poly. added = makes into a double strand which codes for specific protein
DNA probe used to locate a gene = cut out using restriction enzymes

35
Q

alternative ways to obtain gene in genetic engineering

A

nucleotide sequence known - gene synthesised using an automated polynucleotide synthesiser
or PCR primers to amplify the gene

36
Q

explain how genes are placed into a vector in genetic engineering

A

plasmids from bacteria are mixed with restriction enzymes = cuts plasmid at specific recognition sites
cut ends are sticky ends (unpaired)
nucleotide bases are added = complementary to sticky ends of plasmid
DNA ligase catalyses annealing (binding) of gene and cut plasmid

37
Q

alternative way to place gene into vector

A

gene placed into attenuated virus and carry it into host cell

38
Q

explain the ways that vectors are placed into recipient cell

A

heat shock treatment - making walls/ membranes of bacteria more porous
electroporation - high voltage pulse applied to cell to disrupt the membrane
electrofusion - electrical fields used to introduce DNA into cells
Transfection - DNA packaged into bacteriophage , transfects the host cell
gene placed in attenuated v. carried into host cell
gene gun - small pieces of gold used, coated in DNA, fired into plant cells

39
Q

define restriction endonuclease enzymes

A

cut DNA at certain recognition points

40
Q

define vector

A

carry/ insert DNA into host organism

41
Q

define sticky ends

A

single stranded ends of DNA that has been created by endonuclease enzymes

42
Q

DNA ligase

A

joins DNA fragments
joins sugar and phosphate groups of DNA backbone

43
Q

ethical issues with genetic manipulation

A

microorgansims
-escape, antibiotic resistane
- pathogens/virus may be inserted into genome which increases risk of cancer
plants
- super weeds created eg. resistance of herbicides in soya and weeds getting destroyed
- concerns about eating genetically modified food
- no choice whether to but gm seeds
pharming
- using genetically modified animals to make human proteins

module 6 Genetics and ecosystems (3 decks)

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