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gene technologies
By Daniella Di-Fonzo
Specification
(b) outline how gene sequencing allows for genome-wide comparisons between individuals and
between species (HSW7b);
(d) explain that genetic engineering involves the extraction of genes from one organism, or the
manufacture of genes, in order to place them in another organism (often of a different species) such
that the receiving organism expresses the gene product (HSW6a);
(e) describe how sections of DNA containing a desired gene can be extracted from a donor organism
using restriction enzymes;
(f) outline how DNA fragments can be separated by size using electrophoresis (HSW3);
(g) describe how DNA probes can be used to identify fragments containing specific sequences;
(h) outline how the polymerase chain reaction (PCR) can be used to make multiple copies of DNA
fragments;
(i) explain how isolated DNA fragments can be placed in plasmids, with reference to the role of
ligase;
(j)
Sequencing a genome
Genome
Gel electrophoresis
DNA probes
Sequencing a Genome
If
a whole genome needs sequencing: use PCR to make multiple copies of all the DNA.
Squeeze through a tiny hole under high pressure. Cuts DNA into 2000- 10000 base
pairs. These can be shortened to make it easier to sequence them.
Make multiple labelled copies of these lengths of DNA. They are mixed with a primer
(up to 20 base pairs of DNA, complimentary to the start of the DNA fragment you
want), free nucleotides, DNA polymerase (to create new DNA strands) and
nucleotides labelled with fluorescent dye. Each base has its own colour and each
labelled nucleotide with stop the chain from growing.
The DNA polymerase attaches free nucleotides to the lengths of DNA. Most of the
copies wont be complete due to the labelled nucleotides. This means lots of different
length chains.
This is then separated using electrophoresis so the DNA is drawn towards the positive
end of a capillary tube.
A computer records the colours as they pass, the shorter the chain, the faster it
travels, so if there are enough fragments, the computer will be able to work out the
sequence of bases in that particular length of DNA.
This is then done for every piece of DNA and then a computer program works out the
overall sequence of nucleotides in the whole genome.
PCR
PCR can be used to make millions of copies of a DNA
fragment in just a few hours.
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Electrophoresis
This separates different DNA fragments according to their lengths.
A
Electrophoresis (cont.)
Probes form hydrogen
bonds with stretches of
DNA close to its
complimentary
sequence
We
Restriction enzymes
You can get a DNA fragment from a length of DNA using
restriction enzymes.
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DNA ligase
Catalyses
Genetic engineering
Using
technology to change
genetic material of an
organism.
DNA containing lengths from
two different species is called
recombinant DNA
Organisms with altered DNA
are transformed organism
Genes can be manufactured
instead of extracted
An organism with DNA from
another organism is a
transgenic organism
Human insulin
People
with type 1 diabetes need to inject insulin to regulate their blood glucose
concentration
Used to be extracted from pigs, now its mad by genetically engineered bacteria
1. Gene for human insulin is identified and isolated using restriction enzymes
2. A plasmid is cut open using the same restriction enzyme used to isolate the gene
3. The insulin gene is inserted into the plasmid to make recombinant DNA
4. The plasmid is taken up by bacteria and any transformed bacteria are identified
using marker genes
5. The bacteria are grown in a fermenter and human insulin is produced as bacteria
grow and divide
6. The human insulin is extracted and purified
.Advantages
1. More effective than animal insulin and less chance of rejection because its
identical to human insulin
2. Cheaper and faster to produce, more reliable and larger supply of insulin
3. Overcomes ethical or religious issues arising from using animal insulin
Golden rice
Genetically
xenotransplantation
Animals may be used as organ donors for humans when there is a shortage of
human donors
However, there is a risk of rejection
Scientists are trying to eliminate this risk by:
Inserting genes for human cell surface proteins into a newly fertilised animal embryo.
The genes integrate into the animal DNA lowering the risk of rejection
2)
Genes for animal cell surface proteins are inactivated. Removed from the nucleus of
animal cell and then transferred into an unfertilised animal egg. The egg cell is then
stimulated to divide and the animal created doesnt produce animal cell surface
proteins
e.g., a pig that doesnt have the enzymes needed for it to make a sugar that humans dont
have but pigs usually do.
1)
Gene therapy
Somatic alters
the alleles in
body cells.
Offspring can
still inherit the
disease e.g.,
cystic fibrosis
Two types of
gene therapy
Advantages
disadvantages
Dna probes
Can
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Comparing genomes
Comparing genomes of different species