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BMSC 210 > Module 7 > Flashcards

Module 7 Flashcards

1
Q

What is heterologous expression?

A

Expressing a gene in a different host

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2
Q

What is genetic engineering

A

Using in vitro techniques to alter genes in the lab

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3
Q

What is DNA amplification

A

DNA replication in a test tube

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4
Q

How to the cycle numbers grow in PCR ?

A

Each future cycle doubles the number of copies.

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5
Q

What are the PCR steps?

A

I. Denature DNA
2, annealing → oligonucleotides added
3. Add DNA polymerase
4. Heat and cool

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6
Q

What is reverse transcription PCR

A

Makes DNA copies from mRNA template

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7
Q

What does gel electrophoresis do?

A

Separates DNA molecules based on size and charge

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8
Q

What is DNA’s charge?

A

Negative

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9
Q

What is nuclei acid probing

A

Used to detect a certain nucleic acid sequence

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10
Q

How is the probe usually labelled or tagged?

A

With fluorescence

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11
Q

What is a probe?

A

Probes are complementary to the gene of interest

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12
Q

What is a southern blot

A

DNA is in the gol and probe is RNA or DNA

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13
Q

What is a northern blot

A

RNA is in the gel and the probe is DNA or RNA

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14
Q

What is fluorescent in situ hybridization

A

Uses a fluorescent probe attached to oligonucleotide to target specific nucleic acid sequences in cells

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15
Q

What are the enzymes used for cloning?

A

Restriction endonucleases
DNA ligase
Reverse transcription
DNA polymerase

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16
Q

What are restriction endonucleases

A

Enzymes that recognize specific DNA sequences and cut them

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17
Q

What are restriction endonucleases essential for?

A

In vitro DNA manipulation and gene cloning

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18
Q

What is methylation?

A

Coding on cells to prevent cutting by restriction enzymes (defense)

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19
Q

Are restriction endonucleases rare in eukaryotes

A

Yes

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20
Q

What do type 2 restriction endonucleases do?

A

Cleave DNA within recognition sequence: most useful for specific DNA manipulation

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21
Q

What recognizes palindromes

A

Type 2 restriction endonucleases

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22
Q

What is also known as an overhang cut

A

Sticky end cut

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23
Q

What is blunt end cutting

A

Both strands are cut at the same position

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24
Q

When are sticky ends produced?

A

When restriction enzymes make staggered cuts

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25
Q

What can molecules with complimentary sticky ends easily do?

A

Anneal or form hydrogen bonds between complimentary bases

26
Q

What is ecoR1

A

A type 2 restriction enzyme that is isolated from strains of E coli

27
Q

What is each restriction enzyme paired with?

A

A corresponding modification enzyme that shares the same recognition sequence

28
Q

How can a gene to be cloned be amplified by? (3)

A

Polymerase chain reaction
Synthesized by reverse transcriptase
Synthetic DNA made in vitro

29
Q

What are used as cloning vectors

A

Plasmids

30
Q

What are the 3 main steps of gene clothing?

A

Isolation and fragmentation of source DNA
Insertion of DNA fragment into cloning vector
Transformation of cloned DNA into host organism

31
Q

What is blue / white screening

A

Bacteria with a cloning vector inside that may or may not have recombinant DNA

32
Q

What do blue colonies not have

A

Plasma vectors with foreign DNA inserted

33
Q

What do white colonies have

A

The recombinant DNA

34
Q

What gene does blue/ white screening rely on?

A

LacZ gene

35
Q

Where does the lacZ gene often fuse?

A

To a portion of the plasmid known as the multiple cloning site

36
Q

When X gal is cleaved by b-galactosidase, what happens?

A

It turns blue

37
Q

Why does a colony stay white?

A

Because it contains the gene or interest and lacZ gene will be disrupted and b-galactosidase cannot be produced

38
Q

What is on the plasmid vector

A

LacZ gene and a multiple cloning site

39
Q

What are DNA cassettes / cartridges

A

Synthetic fragments that can make more then a few base pair changes or replace sections of a gene via cassette mutagenesis

40
Q

what are the properties of an open reading frame (4)

A

ribosome binding site seuqunce before the start codon
start codon
coding sequence
stop codon

41
Q

what are the enzymes in DNA cloning

A

DNA polymerase
Restriction endonuclease
DNA ligase
Reverse transcriptase

42
Q

whats the purpose of DNA polymerase in cloning

A

it synthesizes the DNA strand

43
Q

what is the purpose of restriction endonuclease in DNA cloning

A

they recognize specific DNA sequences and cut DNA

44
Q

what is the purpose of DNA ligase in cloning

A

going two strands of DNA

45
Q

what is the purpose of reverse transcriptase

A

converts RNA into DNA

46
Q

what do expression vectors have that coning vectors don’t

A

inducible promoter
tag for purification

47
Q

what are expression vectors for?

A

they are designed to facilitate gene expression and protein production

48
Q

what is an inducible promotor?

A

a DNA sequence that drives the expression of the gene - it controls initiation of transcription by RNA polymerase

49
Q

what is a cloning vector?

A

DNA molecule used to carry and replicate foreign DNA fragments in a host organism

50
Q

what is the best generic method to diagnose an RNA virus

A

RT-PCR

51
Q

why is the RT-PCR the best genetic method to diagnose an RNA virus

A

the reverse transcriptase allows the synthesis of cDNA from RNA

52
Q

what is genomics

A

the study of the entire genome (all genes and genetic material)

53
Q

what us transcriptomics

A

the study of a transcriptome, which is the complete set of RNA molecules (mainly mRNA)

54
Q

what is proteomics

A

the study of the proteome, which is the entire set of proteins expressed by the genome

55
Q

what is metabolomics?

A

the study if a metabolome, which is the small molecule metabolites (sugars, lipids, amino acids)

56
Q

what does transcriptomics focus on understanding

A

gene expression patterns

56
Q

what are microarrays

A

they use pre designed probes to measure the expression levels of thousands of genes at once

57
Q

what is western blotting used for

A

measuring protein levels

58
Q

why are primers required for polymerase chain reactions

A

to provide a starting point for DNA polymerase

59
Q

what is a thermocycler

A

a machine used for PCR reactions

60
Q

what is the structure of a nucleic acid probe

A

ss DNA fragments complementary to the sequence of interest

61
Q

cutting DNA with EcoR1 will produce

A

5’ overhangs

BMSC 210 (10 decks)

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