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Biochemistry > Quiz 3 > Flashcards

Quiz 3 Flashcards

(53 cards)

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1
Q

how many Gb is the human genome?

A

3.1 Gb

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2
Q

How many errors

A

around 1 in 100,000

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3
Q

How many variants of genome between individuals?

A

around 4 millions

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4
Q

What percentage of the genome is comprised of segmental duplications?

A

5.3%

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5
Q

How many protein coding genes?

A

19599

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6
Q

How many additional predicted protein-coding genes?

A

2188

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7
Q

How many total protein coding gnes

A

around 22,000

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8
Q

What is the definition of recombinant DNA

A

linking of non-contiguous DNA segments together

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9
Q

In terms of diagnosis what are 4 examples of the function of recombinant DNA

A

predisposition testing
prenatal screening
pathogen id
cancer typing

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10
Q

In terms of prognosis what is the function of recombinant DNA

A

identification of specific mutations

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11
Q

In terms of treatment what are two functions of treatment

A

recombinant proteins

gene therapy

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12
Q

4 clinical uses of recombinant dna

A

diagnosis, prognosis, treatment, forensics

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13
Q

3 experimental uses for recombinant DNA

A

sequencing, protein analyses, gene expression

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14
Q

definition of Restriction Endonucleases

A

sequence-specific DNA binding proteins that catalyze the cleavage of the phosphodiester bonds in dna

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15
Q

How does the restriction endonuclease smaI cut?

A

blunt ends

5-CCC GGG-3

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16
Q

EcoR1 is what kind of base cutter 4,6, or 8

A

6

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17
Q

KpnI is what kind of base cutter 4,6, or 8

A

6

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18
Q

How does EcoR1 cut?

A

5’–G AATTC–3’

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19
Q

How does KpnI cut?

A

5’–GGTAC C–3’

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20
Q

What are the 3 types of DNA polymorphisms

A

Restriction Fragment Length polymorphism
microsatellite repeats
Single Nucleotide Polymorphisms

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21
Q

How are RFLPs detected

A

cutting with restriction enzyme and doing a southern blot

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22
Q

How are microsatellite repeats detected?

Study These Flashcards
A

PCR using flanking primers

23
Q

How are SNPs detected?

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A

pcr followed by hybridization with oligonucleotide

24
Q

What are 4 requirements for PCR

Study These Flashcards
A
  1. DNA sample with known sequence
  2. synthetic oligonucleotides complementary to the target DNA
  3. Thermal stable DNA polymerase (Taq polymerase)
  4. dNTPs
  5. pH buffer and salts for DNA polymerase activity
25
Besides sequence, what is one of the most important features of the primers used for biochem
the primers have to have the correct directionality.
26
Southern blot is for what
DNA
27
Northern blot is for
RNA
28
rt PCR gives you what information
how much mRNA there is
29
What are DNA microarrays
an array of thousands of genes on a very small chip about the size of a postage stamp
30
How are DNA microarrays done?
use PCR to make copies of thousands of different genes, and then use robots to spot a small amount of each of those genes on a very small array
31
Why are DNA Microarrays done?
Examine the expression level of thousands of genes on the chip simultaneously. Compare expression of many genes between 2 different samples
32
Advantage of RNA sequencing | 4
1. Can measure mRNA levels 2. Identifies coding region SNPs 3. Alternative splice isoforms 4. Spurious transcription and unknown genes
33
What are the 3 steps in rtPCR
1. Anneal oligo-dT or other primer to poly (A) sequence at the 3' end 2. Synthesis of first strand cDNA using reverse transcriptase 3. The first strand cDNA becomes the template for PCR
34
can microarray analyses of mRNA measure info on RNA size
no
35
Microarray comparative genomics uses dna or rna?
dna
36
What are the two ways of measuring mRNA levels of specific genes?
quantiative RT-PCR and RNA microarrays/massively parallel sequencing
37
What does microarray comparative genomic hybridization measure?
duplications and deletions
38
How many cells are estimated to be in mitosis at any given time? And how was this estimated?
250 million... surface of epithelial cells and hematopoesis
39
What is the relationship between homologous chromosomes in mitosis?
independent
40
What are the two types of division in meiosis
reductional and equational
41
What happens during reductional division and where is cohesin proteolyzed?
homologous chromosomes were separated and cohesin is only broken down along arms of sister chromatids
42
What happens during equational division and where is cohesin proteolyzed?
sister chromatids are broken down, cohesin is broken down at centromeres
43
Does recombination occur during meiosis 1 or 2
1
44
Differences between mitosis and meiosis
1. Homologous chromosomes are paired up in meiosis 2. 2 rounds of division in meiosis 3. recombination in meiosis
45
What supresses recombination between sister chromatids?
synpatonemal complex
46
4 limitations of Retroviral transduction
1. Silencing of retroviral promoter/enhancer 2. Size limitations ~8kb 3. Common retroviruses only infect dividing cells 4. By integrating into host cell genome, retroviruses have the potential to disrupt important genes
47
Main disadvantage of stable transfections
time consuming b/c ineffective
48
4 requirements for stable transfection
1. regulatory elements 2. gene of interest 3. polyadenylation site 4. separate drug resistance gene with on regulatory elements
49
critical property of ES cells
ES cells can be grown and manipulated in vitro then re-injected into blastocysts to generate embryos= pluripotent
50
knock out
gene disruption or deletion; often results in loss-of-function alteration of a gene
51
knock in
site directed mutation or insertion of new material with the coding region of a gene or its regulatory elements
52
5 steps in producing knockout/knockin
1. create targeted disruption of the gene in ES cell 2. Isolate proper clone 3. Introduce modified ES cells into a 3-1/2 day mouse embryo/blastocyst 4. Generating chimeras from injected blastocysts 5. Breeding chimeras to get "germline transmission" of the modified allele
53
3 disadvantages of hES Cell Use
destruction of embryos rejection tumorigenic

Biochemistry (5 decks)

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