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Psych Final > Bio Test 4 > Flashcards

Bio Test 4 Flashcards

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1
Q

PCR components

A
  • DNA template, gene specific primer pairs, free nucleotides, Taq DNA ploymerase
  • Thermocycle program
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2
Q

94C during PCR

A

opens two strands (denatures) the DNA template

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3
Q

55C during PCR

A

primers anneal (hybridize/connect) to the template, hydrogen bonds begin to form

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4
Q

72C during PCR

A

DNA synthesis (primer extension)

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5
Q

What does PCR do?

A
  • exponential production of specific DNA fragments
  • needs buffer containing Mg+2
  • occurs about 30 times to get many copies
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6
Q

Knock out mice

A
  • removing a genes function

- helicase gene exhibits premature aging

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7
Q

Procedure of knock out mice

A
  • use mouse embryonic stem
  • replace the DNA fragment of interest by recombination
  • mosaic F1 (implant in animal)
  • select for germ-line transmitted animal
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8
Q

genome

A

total genetic information carried by a cell or an organism

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9
Q

transcriptome

A

entire complement of RNAs produced by cell, differential expression comes into play

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10
Q

proteome

A

entire complement of proteins produces by a cell

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11
Q

proteomics

A

systematic study of the amounts, modifications, interactions, localization and functions of all or subsets of proteins at the whole-organisms, tissue, cellular and subcellular levels

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12
Q

characteristics of genomics

A
  • construct genomic and cDNA libraries using recombinant DNA technique
  • cDNAs are the complementary DNAs of mRNAs synthesized by reverse transcriptase enzyme from virus
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13
Q

genomic library

A

combination of recombinant plasmid and E.coli

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14
Q

cDNA

A
  • reversely transcribed from mRNAs by reverse transcriptase enzyme
  • single stranded cDNA can be replicated into double strand cDNA by DNA polymerase
  • build transcriptome
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15
Q

DNA sequencing was developed by who?

A

Frederick Sanger

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16
Q

process of DNA sequencing

A
  • use di-deoxynucleotides to stop the replication
  • only one strand is used as template
  • use one primer instead of two in PCR
  • set up four reactions with regular nucleotides and single dideocynucleotide
  • generate DNA fragments of different lengths
  • run products on gel and read sequence from bottom to top of gel
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17
Q

process of automated DNA sequencing

A
  • use fluorescence labeled dideoxynucleotides
  • reaction is done in one tube
  • data are read and assembled by computer
  • sequencers provide accurate sequences up to 1000 bp long
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18
Q

procedure of genome sequencing project

A
  • generate reocmbinant plasmids containing DNA fragments
  • use automatic sequencer to obtain nucleotide sequences
  • assembly of DNA clones by actual sequences
  • annotation (to identify genes and their locations)
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19
Q

What are the findings of the human genome project?

A
  • human genome contains 3.2 billion bp of DNA
  • 1% actually encodes proteins
  • ~50% DNAs are non-gene sequence repeated thousands of times
  • genes are unequally distributed
  • only have 30000 genes
  • more different proteins than genes alternative splicing 300000 proteins vs 30000 genes
20
Q

What are the four phases of the cell cycle?

A

G1
S
G2
M

21
Q

what is interphase?

A

the collection of phases G1, S, G2

22
Q

what does the G1 phase do?

A

primary growth, longest stage

23
Q

what does the S phase do?

A

DNA synthesis

24
Q

what does the G2 phase do?

A

second growth

25
Q

what does the M phase do?

A

mitosis (nuclear division) and cytokinesis (cytoplasmic division)

26
Q

cell cycle

A
  • has four different phases
  • cells have different cell-cycle times
  • withdrawal from cell cycle: non-dividing cells
27
Q

what does the G0 phase do?

A

resting state

28
Q

cyclins

A
  • expressed in a cyclic fashion
  • no enzyme activity
  • bind and activate Cdks
29
Q

cell cycle checkpoints

A

proteins that check whether call is ready to move on to next phase

30
Q

major cyclins and Cdks of vertebrates

A

S-Cdk complex
M-Cdk complex
G1-Cdk complex

31
Q

S-Cdk complex

A

cyclin A pairs with Cdk2

32
Q

M-Cdk complex

A

cyclin B pairs with Cdk1

33
Q

G1-Cdk complex

A

cyclin D pairs with Cdk4 and 6

34
Q

What are Cdks?

A
  • cyclin-dependent protein kinases
  • activated by cyclins
  • contribute to their own eventual inactivation
35
Q

G1 checkpoint

A

checks whether environment is favorable to begin S phase

36
Q

G2 checkpoint

A

checks for damage and whether DNA is completely replicated

37
Q

mitosis checkpoint

A

checks whether chromosomes are attached to spindles

38
Q

activation of M-Cdk (as an example)

A
  • cyclin B binds and activates Cdk1
  • phosphorylated M-Cdk
  • Cdc25 activates M-Cdk by dephosphorylation
  • positive feedback phosphorylates more Cdc25
  • activate APC
  • complete M phase
39
Q

inactivation of M-Cdk

A
  • APC
  • ubiquitylation of cyclin B
  • degradation of cyclin B
  • inactivation of M-Cdk
40
Q

DNA damage inhibits what?

A

S-Cdk and G1/S-Cdk function

41
Q

process of DNA damage that inhibits S-Cdk and G1/S-Cdk

A
  • DNA damage
  • kinases
  • phosphorylate p53
  • transcription of p21
  • p21 protein binds and inhibits S-Cdk and G1/S-Cdk
42
Q

What are the two types of cell death?

A
  • necrosis

- apoptosis (programmed cell death)

43
Q

necrosis

A
  • osmotic imbalance
  • influx of H2O into cell
  • can burst the cell
44
Q

apoptosis

A
  • carried out by a family of proteases called caspases
  • procaspases are activated through cleavage and reassembly
  • main proteins that regulate the activation of procaspases are members of the Bcl2 family
  • death signal goes to mitochondria first
45
Q

what are the members of the Bcl2 family?

A
  • Bax

- Bak

46
Q

What do Bax and Bak do?

A

form channels on mitochondria causing release of cytochrome C which binds to adaptor protein and then activates first caspase (procaspase 9)

Psych Final (21 decks)

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