[molbio lec] - unit 6: molecular diagnosis of mutation and inherited disorders

chromosome missegregation

mechanism of genome mutation

chromosome rearrangement

mechanism of chromosome mutation

base pair mutation

mechanism of gene mutation

10² per cell division

frequency of genome mutation

6 × 10^-4 per cell division

frequency of chromosome mutation

10^-5 or 10^-6 per locus per generation

frequency of gene mutation

genome mutations

products of the meiotic process

gene mutations

result of environmental factors

true

T/F: genome, chromosome, and gene mutations can all be inherited especially if they occur in the germ line.

genome mutations

most common type of mutation

chromosomal mutation

intermediate type of mutation

gene mutation

rare type of mutation

good DNA repair systems

the reason why gene mutations are rare is because of human’s ____________

achondroplasia

mutation that affects joints

deletion

nature of mutation of Duchenne dystrophy

Hemophilia A and B

examples of a single nucleotide mutation?

Duchenne Muscular Dystrophy

Hemophilia A

Polycystic Kidney Disease (PCKD)

the common mutations within the new/de novo mutations

mutation

A single gene locus causing mutation has a major physiologic impact and considered to be deterministic of a disease

polymorphism

Genetic alteration that contributes to complex disease has smaller effect

polymorphism

present in at least 1% of the population

mutation

seen less commonly in general population

mutation

usually linked to mendelian diseases

polymorphism

mutations that are usually not linked to diseases but alleles are seen more frequently among diseased individuals

mutation

has a one on one correlation with disease

polymorphism

has no one-on-one correlation

point mutations

single base pair change in nucleotide sequence of genes

single nucleotide polymorphisms

single base pair change in nucleotide sequence of genes in polymorphisms

CR5

a mutation/polymorphism involved in the non-entry of HIV-1 into the cell

true

T/F: the distinction between polymorphism and mutation reflects gene function as well as frequency.

environmental factors

if a mutation causes disease or not depends on this particular factor

transition

base pair mutation from pyrimidine to pyrimidine

transversion

base pair mutation from pyrimidine to purine

tautomers

the compounds inside a structure that can have a movement of atoms

amino

a common tautomer

imino

less common tautomer

keto

another common tautomer

enol

less common tautomer

keto

the more stable tautomer

transition mutation

a mutation caused by tautomer of cytosine

replication

the tautomer of cytosine mutation is perpetuated by?

quaternary nitrogen

The formation of a _____________ destabilizes the deoxyriboside bond and the base is released from deoxyribose

escaped radicals

can damage dna and can attack all forms like proteins, lipids, etc.

escaped radicals

in the normal process, they are biproducts of electron transport chain

Oxo DG

a mutation related to escaped radicals

uv light

thymine dimer is formed by what physical factor?

pyrimidine dimers

thymine dimers belong to a larger class of dimers called?

long exposure to UV light

the formation of thymine dimers is due to?

lesions

this means that there is a change in DNA that causes mutation

missing base

acid and heat depurination

altered base

ionizing radiation

alkylating agents

Non-Hemogenous End Joining (HR/NHEJ) pathway

DNA lesion is repaired by?

incorrect base

spontaneous deaminations

deletion-insertion

intercalating reagents

dimer formation

uv irradiation

strand breaks

ionizing radiation

chemicals (bleomycin)

interstrand cross-links

psoralen derivatives

mitomycin c

tautomer formation

fanconi anemia

disease related to interstrand cross-links

wildtype/normal

mos common single base substitution present in the population

nonsense

the transformation is on the stop codon, causing premature protein termination

normal spliced

contains the intron region, and may cause frameshift mutation

tuloy tuloy lang pag splice

exon skipping

magkakadikit yung exon and this splice can create new kinds of proteins that are wrong

can also cause frameshift mutation

hereditary mutations

affects the gametes but does not happen during meiosis itself and rather precedes it

germline mutations

mutations wherein all the cells in the body will experience the mutation

somatic mutation

de novo/new mutations that occur in quickly dividing cells like the skin and the blood cells, usually a result of DNA replication errors

acquired mutations

mutations that are not passed to all the individual cells in the body

point mutations

include missense, silent or nonsense codon changes

affects gene transcript splicing

missense mutations

an alteration in the DNA that results in a different amino acid being incorporated into the structure of a protein

silent mutations

a mutation that still codes for the same amino acid, but can still result in disease

nonsense mutations

a mutation that causes a protein to terminate or end its translation earlier than expected

large or small

deletions and insertions can be either?

frameshift mutations

refers to the insertion or deletion of nucleotide bases in numbers that are not multiples of three

dynamic mutations

unstable trinucleotide repeats

microsatellite expansions and contractions

single-strand conformation polymorphism

scans several-hundred base pairs

based on intra-strand folding

folded single strands can be resolved by size and shape

conformers

folded single strands

allele-specific oligomer hybridization

dot blod method

relies on binding effects of nucleotide mismatches

labeled oligonucleotide probe is hybridized to immobilized specimen

nitrocellulose

specimen in solution in ASO is spotted on?

normal

what does 1 mean in chromosome probe detection?

heterozygous

what does 2 mean in chromosome probe detection?

heterozygous mutant control

what does m mean in chromosome probe detection?

normal control

what does + mean in chromosome probe detection?

negative control

what does n mean in chromosome probe detection?

melt curve analysis

this is based on sequence effect on Tn

can be performed with or without probes

ethidium bromide

sybr green

eva green

three double-strand DNA-specific dyes required for melt curve analysis

fluorescence resonance energy transfer (fret)

this is also performed with melt curve analysis

sybr green

this specific dye will fluoresce when bound to DNA

precision melt analysis

allows differentiation between PCR products based on a single-base pair mismatch

melt curve high resolution analysis

is now used in mutation screening, detection of biological diversity, and genetic analysis

array technology

reverse dot blot methods used to investigate multiple genomic sites simultaneously

substrate

unlabeled probes are bound to?

nitrocellulose

substrate bound to macroarray

glass

nitrocellulose on glass

substrate bound to microarray

glass

substate bound to high-density oligonucleotide

electrode grid

substrate bound to microelectric arrays

microarray

macroarray

array used for comparative genomic hybridization and expression array?

comparative genomic hybridization

Detection of genomic amplifications and deletions

expression array

detection of relative changes in gene expression

snp detection mutation analysis sequencing

Detection of single-base differences in DNA

high density oligonucleotide arrays

Interrogate thousands of genes simultaneously

requires a new array for each sample

tilling

is used to blanket detection of nucleotide changes in the sample

array readers and software

Fluorescence is detected, normalized, and averaged by?