Topic 7 Flashcards
What is a genome
All of an organisms DNA
What is PCR used for
Used to amplify dna by making many copies of a dna sample
What are the steps to PCR
- Reaction mixture is set up by mixing the dna sample , primers , free nucleotides and heat stable dna polymerase
- Mixture heated to 95 degs to break hydrogen bonds and separate dna strands
- Mixture cooled to 55 degs so primers can bind to strands (annealing)
- Temp increased to 70 degs, dna polymerase creates a copy of sample by complimentary base pairing using free nucleotides
- Cycle repeated many times
how can dna sequencing be used to predict the amino acid sequence of proteins
- pcr is involved in amplifying dna to produce many copies of it
- Four tubes set up containing DNA to be sequenced , dna polymerase and primers , terminator nucleotides
A diff chain terminating nucleotide is placed in each tube
DNA strands of diff lengths produced as each one terminates at a diff points
Fragments separated by gel electrophoresis
Fragments move diff distances based on size
Complim base sequence can be read from gel
how can dna sequencing be used to identify possible links to genetically determined conditions?
-some human diseases result from mutations in single genes
through dna sequencing u can identify faulty genes and see which bases have undergon mutations affecting the proteins produced.
whats a terminator base?
modified versions of the 4 nucleotide bases that halt the production of a dna molecule, as soon as theyre incoporated no more bases can be added
What are introns and exons
Introns - non coding dna , the removal of introns is splicing
Extrons - coding dna
What are mini vs micro satellites
As humans we can have the same base sequences but the repeats r diff in each person
Mini-satellites are DNA regions consisting of 10–100 base pair (bp) long sequences that are repeated multiple times.
Micro-satellites are shorter DNA sequences consisting of 1–6 base pairs that are repeated.
How’s DNA profiling done
(used to identify individuals based on their unique DNA sequence. It focuses on analyzing short tandem repeats in non-coding regions of DNA)
- DNA sample extracted and multiple copies made using pcr
- restriction endonuclease enzyme produces dna fragments
- Dna samples placed into buffer solution in wells in agarose gel and an electric current is passed thru. gel electropheresis separates dna according to mass
- Southern blotting - DNA on agarose gel is transferred to nylon filter paper
- Short dna pieces with radioactive markers bind to compllim seq on dna so they can be read by computers
What are transcription factors
proteins that control the process of transcription, a gene is copied into mrna. They decide whether a gene is turned on or off by binding to specific regions of DNA.
Transcription factors bind to specific base sequences through promoter and enhancer sequences
Promoter - enables the binding of rna polymerase and promote transcription, mRNA produced and protein produced which allow cells to undifferentiate
Enhancer regions - binding sites in dna which regulate its activity by changing the structure of chromatin . They can cause dna to be more or less open to rna polymerase. Open =accessible gene available for transcription. Closed = inaccessible and not available
there are 2 ways in which the expression of a gene can be controlled: transcription factors and
splicing of pre mRNA after transcription can result..
in diff products from a single gene
how does RNA splicing work?
- Gene transcribed resulting in pre-mrna
- introns removed and some exons
- spliceosomes join exons many diff ways to produce diff strands mrna for translation
- variations in mrna = code for diff amino acids and thus diff proteins
exons are joined by
enzyme complexes called spliceosomes to produce mature mrna
what is epigenetics
give some examples
-splicing : the way expression of genes can be changed without changing dna base sequ
-dna methylation
histone modification
-non coding rna
-involved in differentiation
histone modification
methylation - addition of methyl group , can cause activation/inactivation of chromatin, linked to silencing genes
acetylation - addition of acetyl group (activates chromatin and allows transcr as it opens up the strcutre)
why are epigenetic modifications important
,they determine whether a gene is switched on or off to ensure cell differentiation, allowing cells to become specialised so they can carry out particular functions
What’s a Closed chromatin structure
Heterochromatin
How can the demethylation of dna cause the conversion of fibroblasts to fuzzy cells
A gene activated
Transcription of gene takes place producing protein that causes change in the cells