BIOL SCI 215 Lecture 16: Lecture 16

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Cancer Genetics
Cancer involves multiple stages - focus on genetics of growth control {alterations to
genomes}
Harvest the DNA from a bunch of tumour cells, compare the DNA sequence and
compare it to the DNA of normal tissue
oDifferent from Sanger sequencing because Sanger is scanning one DNA
sequence
Next generation sequencing
oPerform a reaction similar to cycle sequencing
oLonger read with around 600 base pairs is better for small sequences
oBut if you are interested in many sequences, you will get short reads which
help for open-ended search
Ion torrent technology
oAddition of each dNTP releases a H+ atom, can be detected by transient pH
change
Each target molecule will enter a micron-sized well and the
sequencing will start. Each well has a bead with one DNA clone
When a DNTP is added to a growing strand, a proton is released and
the pH is then measured [H+ release monitored by voltage change”
Line up the reference genome and the sequencer
New mutation is seen by comparison
Tumor cells have many mutations, up to 20,000 per genome
Cancer is an evolutionary process where you have initiating mutations which alter the
ability of DNA to be repaired
oMany steps that the tumor went through to be invasive
Defining genes involved in cancer
oOncogenes are the result of mutations to normal genes that promote cancer
by allowing uncontrollable cell proliferation
oTumor suppressor genes are needed for control of cell proliferation or cell
death after expressive DNA damage
Finding gene alleles that cause cancer “oncogenes”
oTake cells that are isolated from patients and compare them to normal cells
oNormal cells:Fibreglass cells that are connective tissues, plated and grown
into large numbers: they keep dividing and dividing till there is a contact
inhibition phenomenon
oCancer cells keep on dividing and piling up
Cloning on oncogene that is sufficient to transform normal cells into cancer cells
oChop up DNA genome of cancer cells, clone into expression plasmid and
transfect mixture into normal cells
oA few normal cells in dish turn into cancerous cells. Then determine gene that
was transfected into these cells
oCollect DNA cells from cancer cells, cut with restriction enzymes, clone into
expression vector which must have an ori, AmpR because bacteria, and MCS
as well as a mammalian promoter. Put into normal cells by transfecting it.
oRas oncogene expression was enough to take a normal cell and changing it
into cancer cells
Growth factor signaling through receptor tyrosine kinases
oSecreted growth factor produced from distant cell and the signal is received
from cell that is bind by a receptor which causes it to dimerize and activate.
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Document Summary

Cancer involves multiple stages - focus on genetics of growth control {alterations to genomes} Harvest the dna from a bunch of tumour cells, compare the dna sequence and compare it to the dna of normal tissue: different from sanger sequencing because sanger is scanning one dna sequence. Ion torrent technology: addition of each dntp releases a h+ atom, can be detected by transient ph change. Each target molecule will enter a micron-sized well and the sequencing will start. Each well has a bead with one dna clone. When a dntp is added to a growing strand, a proton is released and the ph is then measured [h+ release monitored by voltage change . Line up the reference genome and the sequencer. Tumor cells have many mutations, up to 20,000 per genome. Cancer is an evolutionary process where you have initiating mutations which alter the ability of dna to be repaired: many steps that the tumor went through to be invasive.

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