quiz 3.4 mutation, oncogenes Flashcards
kinds of point mutation
synonymous
missense
nonsense
point mutation can be caused by
insertion of transposon
abberant recombination
repreats of simple tri- di- or tetranucleotide sequences
DNA microsatellites
imino form of cytosine pairs with
adenine
enol form of thymine pairs with
guanine
imino form of adenine pairs with
cytosine
enol form of guanine pairs with
thymine
mutation of purine to purine or pyri to pyri
transition
mutation of pyri to purine or vice versa
transversion
mutation where altered codon corresponds to stop codon
nonsense
depurination leads to
apurinic site
depurination is repaired by
base excision repair
occurs when bases become spontaneously ionized
mispair
thymine analogue that has bromine at C5 instead of methyl group
5-bromouracil
causes G-C to A-T or vice versa transitions
5-BU
keto form of 5-BU pairs with
adenine
ionized form of 5-BU pairs with
guanine
base analog of adenine
2-aminopurine
protonated 2-AP can mispair with
C
rate of spontaneous mutations
10-6 to 10-11 per round of replication
in streisinger model, when template strand slips, ____ results
deletion
in streisinger model, when primer slips, ___ results
+1 insertion
steps in restriction mapping
restriction endonucleases
separation
visulaization
restriction endonucleases commonly cleave
palindromic segments
selectively cut the DNA in a sequence-specific manner
restriction endonuclease
presence of restriction endonucleaserestrict the growth of what
bacteriophage
type of RE commonly used
type 2
generated when cleavage is asymm
sticky/chosive/overlapping ends
generated when cleavage is symm
blunt ends
rest. fragments can base pair with each other if
sticky ends are complementary
two unrelated fragments can base pair with each other if
cleaved by same RE
the ends are linked by
DNA ligase
blunt ends are only linked by
T4 ligase
ligation of two DNA from differetn sources creates a
recombinant or chimeric DNA
gel for higher resolving power
polyacrylamide
more proous, resolves mixtures of larger fragments
agarose
detects specific DNA fragments among thousands of molecules
southern blot
detects RNA fragments instead of DNA
northern blot
detects proteins, used with southern blot
western blot
blotting to study protein-DNA interactions
southwestern blot
steps for souther blot
separate
laying of gel onto nitrocelulose
labeling
reading
methods for determining DNA structure and relation to gene expression
Maxam and Gilbert (chemical)
Sanger (manual enzymatic)
in Maxam and Gilbert, ssDNA is 32P-labeled where
5’-hydroxy end
Maxam and Gilbert uses what separation technique
PAGE
Maxan and Gilbert can sequence stretches of DNA containing around how many bases
250 bases
procedure of choice for DNA sequencing
Sanger
Sanger can sequence up to how many bases
400
Sanger employs what
controlled interruption of enzymatic replication
in Sanger, this required to introduce segment into host cell where complementary DNA is synthesized
vector
______ of each base is mixed with 4 dNTPs together with e. coli DNA polymerase
2,3-dideoxynucleotide
dNTP when incorporated into the growing DNA strand causes what
strand to stop
commonly used cloning vectors
plasmid bacteriophage lambda cosmids ssBacteriohage vector expresson vector
genes within a plasmid
for antibiotic resistance
restriction sites (cloning sites)
origin of replication
double stranded, closed mini-circular extra-chromosomal DNA containing 1-200 kbp, capable of independent replication
plasmid
plasmid occurs naturally in some bacteria as
accessory chromosomes
can destroy its host or become part of its host, bacterial virus containing a double-stranded linear DNA
bacter lambda
ideal cloning vectors for DNA up to 20 kbp, usually for eukaryotic
bacte lambda
plasmids that contain DNA sequences from baceriophage with combined features of plasmids and phages
cosmids