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BIOC3021 > Recombinant DNA > Flashcards

Recombinant DNA Flashcards

1
Q

____ is producing exact copies of a DNA segment

A

cloning

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2
Q

_____ _____ is the ____ _____ in which foreign DNA is inserted for cloning

A

cloning vector, DNA molecule

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3
Q

What are the 4 common types of cloning vectors?

A

plasmids, cosmids, bacterial artificial chromosomes (BAC), and yeast artificial chromosomes (YACS)

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4
Q

Cloning vectors with their inserted foreign DNA can be introduced into host cells where they are able to _____

A

multiply

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5
Q

Is cloning in vitro on in vivo?

A

in vivo

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6
Q

Is PCR in vitro or in vivo?

A

in vitro

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7
Q

In vivo means occurring where?

A

in a host cell

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8
Q

In vitro means occurring where?

A

in a test tube

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9
Q

What cloning vectors can fit the largest size of DNA insert?

A

YAC

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10
Q

What cloning vector fits the smallest size of DNA insert?

A

plasmid

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11
Q

Foreign DNA fragments are inserted into the ____ ____ ______

A

plasmid cloning vectors

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12
Q

The plasmid vector and foreign DNA are treated with a _____ _______

A

restriction endonuclease

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13
Q

_____ ______ is the enzyme that cuts DNA at a specific sequence

A

restriction endonuclease

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14
Q

Restriction endonuclease creates ____-______ ends when cutting

A

single-stranded

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15
Q

Single-stranded ends are useful for what two purposes in cloning?

A

base-pairing of the foreign DNA and plasmid vector, resealing the backbone by linking the backbone of the foreign DNA and plasmid (a covalent linkage)

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16
Q

_____ is the term for the base-pairing of the single-stranded ends

A

annealing

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17
Q

Ligating is the _____ linking of the backbone of the foreign DNA and the plasmid vector

A

covalent

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18
Q

The recombinant DNA has a _____ circumference than the original plasmid

A

larger

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19
Q

The base-pairs of the foreign DNA ____ bond with the base pairs of the plasmid vector

A

Hydrogen

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20
Q

A _____ plasmid is one with partly foreign DNA And partly plasmid SNA

A

chimeric

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21
Q

The DNA backbones of the plasmid vector and the foreign DNA ____ link with each other

A

covalently

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22
Q

A ____ _____ is a set of cloned chromosomal DNA fragments representing the entire genome of an organism

A

genomic library

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23
Q

Isolating DNA, treating DNA with a specific enzyme to be cut into fragments, producing fragments 4,000 bp in length, and placing the fragments into unique plasmids is the process used to produce ____ _____

A

genome libraries

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24
Q

A ____ _____ is a set of cloned cDNAs representing all of the mRNA (gene transcripts) synthesized by cells or tissues

A

cDNA library

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25
Q

cDNA is useful for ____ ____ because it does not have ____

A

protein expression, introns

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26
Q

______ are DNA copies of mRNA molecules

A

cDNAS

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27
Q

The primer connects to the ________when synthesizing cDNA molecules

A

poly-A tail

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28
Q

____ ______ uses RNA sequences to copy into DNA and adds new DNA nucleotides during cDNA synthesis

A

reverse transcriptase

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29
Q

The heteroduplex is the hybrid ___/____ molecule

A

DNA RNA

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30
Q

_______ attacks RNA to remove it when making cDNA

A

RNase H

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31
Q

___ ______ copies first strand cDNA using RNA segments as primer

A

DNA polymerase

32
Q

When making cDNA, DNA fragments are joined by ___ _____ to make the end helix

A

DNA ligase

33
Q

A vector for in vitro expression of a cDNA molecule or other foreign DNA can be constructed by placing a ____ _____ next to the ___ _____

A

promoter sequence, inserted DNA

34
Q

In RNA expression, the host ___ _______ bonds to the plasmid with foreign DNA to produce mRNA

A

RNA polymerase

35
Q

To express a protein in a host cell, a cDNA molecules is inserted next to a ___ _____ and a _____-____ ______

A

promoter sequence, ribosome-binding site

36
Q

A _____ ______ ______ is a sequence of 6 restriction enzymes that are the recognition and cutting sites and is where you cleave the plasmid to insert DNA

A

polylinker cloning site

37
Q

The mRNA produced via protein expression vector contains the ____-____ _____ but not the promoter

A

ribosome-binding site

38
Q

The benefit of producing protein via protein expression vector is that cells can later be harvested and the proteins purified and what?

A

cells can be frozen and stored to be used later to restart protein synthesis

39
Q

PCR is used to _____ a specific _____ ______

A

amplify, DNA segment

40
Q

The 3 steps to PCR are ____, ____, and _____

A

separating, annealing, extending

41
Q

PCR ______ occurs at the highest temp

A

seperating

42
Q

PCR primers are ____

A

oligonucleotides

43
Q

The target DNA for PCR does/does not need to be purified

A

does not

44
Q

The amount of DNA _____ with each cycle of PCR

A

doubles

45
Q

DNA polymerase are heat _____

A

sensitive

46
Q

What is the DNA polymerase that is heat sensitive and used in PCR?

A

taq polymerase

47
Q

___ ______ eliminates the need for fresh DNA polymerase for each cycle of PCR

A

Taq polymerase

48
Q

___ ______ cut DNA at specific sequences and have subsequently been used for cloning and sequencing of DNA molecules

A

restriction endonucleases

49
Q

If the res. enzyme cuts an even number sequence, they are _____

A

palendromes

50
Q

Res. enzymes that make staggered cuts produce _____-_____ ends

A

single strand

51
Q

Res enzymes that make _____ cuts produce single-stranded ends

A

staggered

52
Q

Res enzymes that make non-staggered cuts produce ____ ends

A

blunt

53
Q

Rest enzymes that make ____-____ cuts produce blunt ends

A

non-staggered

54
Q

_____ end cuts are less useful in cloning

A

blunt

55
Q

_____ cuts go down the middle

A

blunt

56
Q

___ ______ is the blotting method where DNA fragments are separated by gel electro., transferred to a sheet of membrane, and detected with a probe

A

southern blotting

57
Q

___ ____ is the blotting method where RNA molecules (such as mRNAs) are separated in a gel, transferred to a sheet of membrane, and detected with a probe

A

northern blotting

58
Q

____ _____ is the blotting method where proteins are separated in a gel, transferred to a sheet of membrane, and detected with antibodies

A

western blotting

59
Q

Southern blotting is used for ____ detection

A

DNA

60
Q

Northern blotting is used for _____ detection

A

RNA

61
Q

Western blotting is used for ____ detection

A

protein

62
Q

If the desired protein is detected in western blotting, it will bind to the _____

A

antibodies

63
Q

____ ____ ______ is the introduction of a functional gene into cells of human tissues and organs to correct damage caused by a mutant gene

A

human gene therapy

64
Q

Human gene therapy is used to produce _____ _____ in ___ ______

A

human protein, human cells

65
Q

An ___ ______ consists of a cDNA version of a gene and a promoter for human gene therapy

A

expression cassette

66
Q

In gene therapy, the expression cassette is incorporated into a _____ that can infect human cells

A

virus

67
Q

Viral vectors for human gene therapy include ____ ____ ____ and _____ ______

A

DNA virus vectors, retrovirus vectors

68
Q

Human gene therapy occurs in vivo or in vitro?

A

in vivo

69
Q

The DNA virus used in human gene therapy is the _______

A

adenovirus

70
Q

Adenovirus vectors are produced by deleted __ ____ to create space for the ___ _____

A

gene E1, expression cassette

71
Q

The complementing cell line contains the E1 gene which allows the adenovirus vector to ____ and become _____ into infectious virus

A

replicate, incorporated

72
Q

The adenovirus from the complementing cell line is used to ____ the cells of a patient and express the ___ _____

A

infect, functional protein

73
Q

Cystic fibrosis, sickle-cell, hemophilia, and muscular dystrophy are examples of what type of disease?

A

single-gene

74
Q

Rank the sizes of the DNA cloning vectors: YAC, plasmid, BAC, cosmid

A

plasmid<cosmid<BAC<YAC

75
Q

cDNAs are DNA copies of _____ molecules

A

mRNA

76
Q

The 3 steps to PCR are ____, ____, and ____

A

separate, anneal, extend

77
Q

In PCR, what component of the reaction mixture is altered to introduce a mutation into the nucleotide sequence of a gene that is to be amplified?

A

primers

BIOC3021 (25 decks)

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