Mutations Flashcards
Explain why most mutations on the third position of a codon results in a silent mutation
- Multiple codons can code for the same amino acid
- Change in the third position on codon would in most cases still code for the same amino acid e.g. GUG -> GUA would be a silent mutation as they both code for Valine
Name the 4 types of mutations that may affect the codons coding for amino acids
- Missense
- Nonsense
- Silent
- Frameshift
What is meant by a MISSENSE mutation?
Single base change that results in a substitution of amino acids e.g. GGC -> TGC substitutes Gly -> Cys
What is meant by a NONSENSE mutation?
Single base mutation that results in a codon coding for an amino acid becoming a stop codon e.g. GGA -> TGA substitutes Gly -> STOP
What is meant by a SILENT mutation?
Single base mutation that does not result in an amino acid change e.g. GUG -> GUA still codes for VALINE
Define a FRAMESHIFT mutation and state the consequences
- Base insertion or deletion that results in the alteration of the mRNA reading frame
- mRNA may be destroyed or code for a different protein
- Insertions, deletions or splice site mutations
- Creation of a PREMATURE TERMINATION CODON
What are the consequences of a splice site mutation?
- Results in the corresponding EXON being cut out of the reading frame
- EXON would not be coded for
- Mature mRNA reading frame would be different (mRNA may be degraded)
Explain how a missense mutation may NOT have a great effect on the overall protein structure and function
- Amino acid substitution results in an amino acid change to one of a similar chemical nature
- For example GTN -> GCN would result in an amino acid change of Val -> Ala which are both non polar and hydrophobic
- Resulting protein will be very similar
What is the difference between a transition and a transversion?
- TRANSITION is a base change to a base of the same type e.g. purine —> purine
- TRANSVERSION is a base change to a base of a different kind e.g. purine —> pyrimidine
Describe how a single base mutation can be detected using various genetic tests
- PCR amplification of DNA fragment
- Hybridisation with ALLELE SPECIFIC OLIGONUCLEOTIDE
- DNA sequencing
Explain when you would use an allele-specific PCR method to investigate a mutation
- For known-disease causing mutations
- Two different allele specific probes have one common primer
- Primer is perfectly complementary to ONE of the probes (one will bind less tightly as it is the same apart from a single base)
- This can be either the probe complementary to the normal gene or the probe complementary to the mutated gene
- PCR primer will only amplify if the allele specific probe matches perfectly to the primer
When would you use Southern blotting over allele specific PCR when investigating a mutation?
- With partial gene inversions the gross organisation needs to be investigated
- Southern blotting allows investigation of an individual gene in a background of all other genes
- Analysis of larger segments with or around the gene
- Can be used to analyse triplet repeat/deletion disorders and resulting fragments may be of different sizes
What is the most common base pair transition in the human genome?
C to T
Explain why the genetic code can be described as both degenerate and unambiguous
Each codon codes for one specific amino acid (ambiguous) but each amino acid can be coded for by multiple codons (degenerate)
How would a deletion of 3bp affect the reading frame of the mRNA strand?
- Shorter mRNA strand produced
- Reading frame would remain intact
- Would result in loss of amino acid therefore a shorter protein
Name 3 occurrences which can result in base changes of DNA
- Sequence changes during DNA replication
- Chemical induced mutations
- Radiation exposure
What 2 spontaneous occurrences can cause base changes during DNA replication?
- Tautomeric shift
- DNA strand slippage
Describe how tautomeric shift can result in a base change of DNA
- Proton briefly changes position on base forming a rare form which has different H bonding properties
- Rare tautomeric forms cause C to bond with A and T to bond with G
- DNA polymerase reads base in tautomeric form and bonds an incorrect base into the new strand
