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Molec > Exam 1 > Flashcards

Exam 1 Flashcards

1
Q

Muller

A
  • -> mixed water, hydrogen, methane, carbon dioxide, ammonia through an apparatus with electrical spark to get amino acids
  • ->Tested Radioactive Particles affect on genes, test on male fruit flies (exposed them to relatively high doses of radioactivity) induced over 100 mutations resulting in progeny
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2
Q

Griffith

A

Studied bacteria that causes pneumonia

  • -> rough strain is non pathogenic
  • -> smooth strain is pathogenic
  • –> heat killed S and R causing mice to die
  • -> transformation : allows the take up of DNA
  • R cells took up the DNA so it was able to make the capsules
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3
Q

McCarthy, Avery & Maclead

A

Added treatment that destroyed RNA ad proteins that are In little amounts of DNA, transformation occurred. Then treatments that killed DNA stopped transformation of S cell DNA to R cell DNA completely
DNA carries genetic info not proteins
–> but people still didn’t believe this

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4
Q

Hershey and Chase

A

proved DNA carries genetic info
–> phage DNA directs the reproduction of virus in infected bacterial cells by it entering the bacteria after attachment but the protein coat or virus remains outside

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5
Q

Franklin

A

produced an extra image of DNA the pattern represent the helix shape

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6
Q

Watson & Crick

A

used Chargaff’s rule and franklin image to make the double helix model

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7
Q

Chargaff’s Rule

A

purine + pyrimidine
T:A
C:G

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8
Q

Central Dogma of life

A

implies there is amplification

  • -> the host DNA replication machinery makes copes of recombinant DNA (two species DNA put together)
  • -> RNA polymerase make multiple mRNAs
  • -> Each mRNA directs many ribosomes to synthesis protein
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9
Q

Understanding the central dogma of life

A

bacteria can be directed to overproduce almost any protein whose gene can be isolated

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10
Q

Beadle & Tatum

A

Selection and complementation of microorganism builds the one enzyme one gene hypothesis to dissect biochemical pathways
–> inborn earros of metabolism can be studied as genetic markers
–> induced mutations in microorganisms more efficient
-mutations in different genes are complementary
the ability to grew on pathway intermediates can establish the order in which enzyme from the genes act

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11
Q

Biochemistry

A

is a result of evolution from a single ancestor

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12
Q

Bases: purines

A

adenine, guanine, hypoxanine

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13
Q

Bases: pyrimidine

A

cytosine, thymine, uracil

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14
Q

Parts of DNA and RNA

A

DNA or RNA are the backbone
3’ to 5’
hydrogen bond donor (Nh2) and acceptor (O)

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15
Q

Parts of a chromosome

A
  • two subunits called chromatids
  • Kinetochore: complex of DNA and proteins to which the spindle fiber attach
  • Euchromatin: expressed part
  • heterochromatin: not expressed
  • telomeres: caps
  • centromeres - holds the chromosome in the middle
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16
Q

Interphase

Checkpoints of interphase

A

G1: gathers proteins and materials to prepare for the S phase; no DNA replications
S phase: DNA replication
G2: preparing organelles and components to enter mitosis; post DNA synthesis; no DNA replication
–> attempts to prevent mutated cells from dividing
P53 can stop the cell cycle and cause the cell to die

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17
Q

Mitosis

A

dividing of autosomes

2n–> 2n

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18
Q

Prophase

A

condensation of chromosomes
nucleoli disappears and nuclear envelope disintegrates
- in meiosis this is where cross over occurs

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19
Q

Metaphase

A

spindles form (consists of microtubules formed by tubulin)
–> elongates out of centrosomes
–> centromeres are duplicated during interphase before mitosis
kinetochore - where spindles attaches to the chromosomes centromeres; kinetochores lie equidistant from the spindle poles
–> each chromatid is regarded as a separate chromosome; align on the metaphase plate

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20
Q

Anaphase

A

proteins holding the centromeres together are degraded
two sister chromatids move to opposite ends of the spindle poles
–> now each considered a chromosome
each group on either side consists of the same number of chromosomes that the original cell had during interphase

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21
Q

Telophase

A
  • ->nuclear envelope forms around group of chromosomes and the spindle disappears and chromosomes decondense
  • -> nucleoli are formed
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22
Q

cytokinesis

A

formation of cleavage furrow
actin rig form tw separate membranes
seals off the cell to make two daughter cells

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23
Q

Meiosis

A

sex cells (2n–>n) with unique gametes

  • -> meiosis 1: generates 2 haploids cells (separation of homologous chromosomes)
  • -> meiosis 2: generates 4 haploids cells *separation of sister chromatids)
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24
Q

Difference Mitosis and Meiosis

A

b/t individual pairs of homologous chromosomes but sister chromatids are also different
–> chiasma - cross over point

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25
Q

Chromatin

A

DNA wrapped into nucleosomes

- nucleosomes: two coils of double helical DNA wrapped around a protein spool

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26
Q

Protein parts

A
  • an octamer of histone proteins make the core
  • core acts as a spool about which DNA winds
  • histone: DNA interaction are non specific - any DNA sequences can be wrapped into nucleosomes
  • -> histone 1 is outside of core binds linker DNA b/t neucleosome, only one with only one
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27
Q

DNA part

A
  • 140 bp of DNA make just under two tight wraps about the octamer core
  • a linker DNA connects the wrapped spools (straight)
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28
Q

DNA is condensed by coiling

A

nucleosomes pack into coil-filaments

  • 30 nm diameter
  • can’t easily be transcribed
  • not in mitosis in this size
  • organized into domains
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29
Q

DNA condenses

A

filaments get coiled repeatedly

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30
Q

Special enzymes modify chromatin to control

A
  • gene access and transcription
  • histones are modified to loosen us
  • -> filaments and DNA interaction
  • DNA itself can be modified to give relative effects
31
Q

Complementation Test

A

bring two mutant genes together in the same cell or organism
–>#of complementations groups= # enzymes

32
Q

Non mutant phenotype

A

are said to complement one another and it means the mutations are in different genes

33
Q

mutant

A

fails to compliment one another and it means the mutations are on the same gene so in the same complimentation group

34
Q

Matrixes

A

+ means that the indicated mutations do compliment on another

  • means that the indicated mutations don’t compliment on another
  • -> if something doesn’t compliment with anything else it is considered its own gene
35
Q

Phenotype and genotype

A

phenotype: what you see
genotype: allele
dominant: expressed
Recessive: masked

36
Q

Codominant

A

phenotype of both alleles expressed –> blood type

heterogeneous genotype has traits associated with both homologous genotype

37
Q

incomplete dominance

A

a cross between organism with two different phenotype produces offspring with third phenotype that is a blending of the parental traits
- intermediate trait of both the dominant and recessive

38
Q

Rules of probability: addition rule

A

the probability of the realization of one or the other of two mutually exclusive events, A OR B, the sum of their separate probabilities

39
Q

Rules of probability: multiplication rule

A

the probability of two independent events, A AND/WITH/BOTH B, being realized simultaneously is given by the product of their separate probabilities

40
Q

Testcrosses

A

used to determine the genotype of F1
cross dominant phenotype (unknown genotype with recessive phenotype
example: progeny you get is half round and half wrinkles, 1:1 phenotype it could have been all round or all wrinkled, so the genotype of F1 is heterozygous

41
Q

Backcrosses

A

determine the genotype of hybrid
cross hybrid genotype with parental genotype (known)
resulting genotype is closer to the parent

42
Q

Expressivity and Penetrance

A

Expressivity: genes are expressed to different degree in different organisms

  • phenotype may vary in expression of particular genotype because
  • -> other genes modify the phenotype
  • ->that phenotype is sensitive to environment
    penetrance: refers to the proportion of organism who phenotype matches their genotype for a given trait. a genotype that is always expressed has penetrace to 100%
43
Q

Epistasis

A

need genes to get the required phenotype, the genes code for enzymes in the same biochemical pathway
–> masking

44
Q

polygenic trait

A

more than one gene responsible for the trait

–>eye and skin color

45
Q

pedigrees

A

dominant doesn’t skip generation
recessive can skip generation
inbreeding results in an excess of homozygotes compared to normal mating - rare recessive alleles, that wouldn’t otherwise become homozygous appear

46
Q

X-linked diseases

A

more commong in males than females because males only have one X and women have 2
women are carriers
Hemophila A
Color Blindness

47
Q

wm/++

A

means the mutant alleles are on the same chromosome and the wildtype alleles are on the same chromosome (cis)

48
Q

Progeny

A

has the same phenotype as on of the parents then it is non recombinant
has phenotype is a mix of parents phenotype together then it is recombinant

49
Q

Recombination frequency

A

total number of recombination (non-parental) divided by total # of progeny (including parent

  • -> only on x chromosome
  • -> different from one gene pair to another
  • -> genes located further away are more like to have recombination
  • -> cis: same sister chromosome
  • -> trans: different sister chromosome
  • -> if it is smaller than 50% than the genes are on the same chromosome (linked)
  • -> 50% recombination (the max) means independent assortment and the genes are either on nonhomologous chromosomes or far apart on the same chromosome
50
Q

Chromosome interference

A

crossover in one region reduces the probability of a second crossover nearby
I=1-(c), coefficient of coincidence: the ratio of observed to expected double recombinants
–> frequency of recombination of total progeny: expected (observed/expected = c)
–> interferences increases with distance

51
Q

Three Point Cross order

A

this will give three genes and give their distances

  • -> reorder genes so same letter is in the middle, so it clearer where the recombination occurs
  • -> parent the two with the highest number
  • -> double recombination the two with lowest number
52
Q

Genetic vs physical map

A

map distance (cM) based on recombination frequencies are not a direct measurement of physical distance along chromosome

  • -> recombination “hot spots” overestimate physical length
  • -> low rates of recombination in heterochromatin and centromeres underestimate actual physical length
  • -> a chromosomal map based on recombination frequencies may be up to 200 cM, but you can’t have 200% recombination
  • -> hot spots( by HATS) are sensitive to endonucleases and that’s why there’s cleavage and that’s how you get recombination
53
Q

Restriction enzymes and DNA ligases cut and paste

A

endonucleases cut palindromic sequences in nucleic acids

  • -> restriction endonucleases: cleave double stranded DNA molecules wherever a particular, short sequences of bases is present –> selective in where they cut the DNA
  • -> DNA ligases make a new phosphodiester bond between two fragments
54
Q

Electrophoresis

A

can be native or denaturing

- larger band stay on top because they are slow and the smaller bands are faster and go towards the + end

55
Q

Gene Cloning: Vector

A

Vector: is a DNA molecule that is able to replicate inside a cell and carries foreign genetic material
–> when a DNA fragment has been joined to the vector, the recombinant molecule is introduced into a cell by means of DNA transformation
–> an isolated transformant containg the recombinant molecule is said to be cloned
key components:
- all vectors are plasmids but not all plasmids are vectors
- vector is a plasmid that is engineered to make it more useful

56
Q

Library construction

A
  • isolate mRNA from a cell
    make the cDNA from the RNA using reverse transcriptase
  • use DNA polymerase to synthesize second strand and PCR to add restriction sites
  • find the sequence of DNA for an organism
  • ligate DNA into vector and introduce into bacteria
  • recombinant DNA - combinding DNa from a couple sources
  • each vector in a library carries a different random piece of DNA
  • find the gene of interest
57
Q

Transformation

A
  • direct uptake of plasmid DNA
  • -> plasmid = circular double stranded DNA (not part of genome)
  • -> RNA polymerase converts DNA to RNA and it binds at the promoter and it stops at the terminator
  • -> conjugation : two bacteria form a conjugal junction and the one with the plasmid can transfer it to the other bacteria and it will duplicate and grow
  • can also be used to get DNA into a cell
58
Q

Bacterial Transformation

A
  • shock bacteria to create small holes in the lipid bilayer (extreme cold) to allow plasmid to go through the pores and its transformed if plasmid are in the bacteria
  • heat bacteria up against so the plasmid stick in the bacteria instead of passing thought the pores
  • no plasmid = no transformed
  • if you put the pacteria on a dish with penicilin and it doesn’t have the plasmid then it won’t grow because it isn’t antibiotic resistant gene from the plasmid
59
Q

Electroporation

A

mix plasmid and cultured cell into cuvette and apply voltage so there’s holds in the cell membrane for the plasma to enter through

60
Q

Transfection

A

mix DNA with lipid solution so the DNa is encapsulated in a lipid bilayer (called liposome)
- liposome can fuse with the cell membrane of a cell so DNA enters the cell and its trasported into the nucleus

61
Q

Transduction

A

use viruses to get DNA or RNA into cell

62
Q

Gene gun

A

plant cells
plastic bulletes shot into ridge cell wall
plasmids are put onto micro particles which are shot into the cell

63
Q

Chromosomal vs cDNA

A

lkjasdlfkj

64
Q

DNA hybridization: probes

A

are solutions of denatured DNA

  • -> they are mixed with genomic DNA fragments and renaturation take place
  • — DNA fragments are able to hybridize only if the length of the region in which they can pair is sufficiently long
  • -> bag is heated causes the DNA to anneal
65
Q

DNA hybridization: Southern blots

A

electrophoresis gel that has separated DNA is placed in contact with a sheet of nitrocellulose so that the DNA is transferred with its position of the DNA brands maintained (through capillary action)
–> only binds to single stranded DNA so the DNA needs to be denatured
the filter paper is put into the heat sealed bag - DNa annealing with probe in solution
take out filter and wash off the membrane because some probe isn’t bound and needs to be removed
fragments start to appear under x ray film

66
Q

PCR

A
  • method for selective replication
  • uses DNA polymerase and short oligonucleoties (as a primer) that are complementary in sequence to the ends of the DNA sequence to be amplified
  • denature DNA sample to separate DNA strands
  • Primers anneal to DNA strands
  • Taq polymerase synthesizes new DNA strands (extends primers)
  • denature to separate strands (cycle)
  • 2^number of rounds
67
Q

DNA Sequencing

A
  • determines the order of bases in DNA
  • sanger method: dideoxysequencing method emplys DNA synthesis in the presence of small amounts of fluorescently labeled nucleotides that contain the sugar dideoxyribose (lack 3’ Oh) instead of deoxyribose to act as a terminator
  • process: put single stranded DNA, primer, DNa polymerase, and the 4 deoxy-bases each in a tube, add small amounts of each didoxynucleotide to their respective tubes; the primers will make strands until a dd base is used which causes it to stop
  • put each tube in electrophoresis –> oligonu are separated by size and nucleotide type;
  • -> read from the bottom to the top
68
Q

Microarray

A
  • mRNA from both not infected and infected
  • use revere transcriptase to convert to cDNA
  • on the DNA chip there are probes of interest (which has the genome on it)
  • probes can hybridize the DNA to make it bind to the chip
  • based on the color you can tell if the DNa being experimented with contains a certain genes
69
Q

DNA polymorphisms: SNPs

A

single nucleotide polymorphism

- single base pair difference

70
Q

DNA polymorphisms: RFLPs

A

restriction fragment length polymorphism

  • electrophoresis, southern blotting, the probes will bind to fragments of interest
  • 1 band: mutant
  • 2 bands : normal
  • 3 bands means carrier, one normal allele, one with RFLP
71
Q

DNA polymorphisms: SSRs

A 
simple sequence repeat 
and STR: short tandem repeats 
- widespread in most naural poluations 
- pedigrees and electrophoreisis 
- see 1 band meand they are on the same exact strand
72
Q

Polymorphism in gernal

A
  • use microarroy to determine which polymorphism a person has
73
Q

Dot Blot

A

immobilized
like southern blotting
use when there is many samples

74
Q

Reverse Blot

A

probe is immobilized

  • good when you many many probes
  • labeled sample in solution

Molec (1 decks)

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