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Molecular Genetics Flashcards

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1
Q

the use of molecular genetics revolution techniques to develop mew products is called

A

Biotechnology

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2
Q

a set of molecular techniques for locating, altering, recombining and studying specific pieces of DNA is called

A

Recombinant DNA

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3
Q

an example of restructuin enzymes

A

site-specific endonucleases

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4
Q

what method are designed to cleave DNA at predetermined sequences

A

Engineered nucleases

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5
Q

what is a facile method for ediiting genomes across the phylogenetic spectrum

A

CRISPR-Cas genome editing

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6
Q

locating DNA fragments can be done with

A

hybridization probes

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7
Q

what enzymes are responsible for recognizing and cutting DNA at specific nucleotide sequences

A

Restriction enzymes

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8
Q

what is the most useful class of restriction enzymes

A

Type II

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9
Q

Restriction enzymes are part of _________ restriction modification system which is a defense against viruses

A

bacterium

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10
Q

each restructuin enzyme is paired with which enzyme that methylates the recognition sequence

A

restriction methylase

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11
Q

function of methylation of recognition sequence

A

blocks cleavage by the restriction enzyme to protect the bacterium’s own DNA from being digested

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12
Q

what does the first 3 letters of the abbreviation for each restriction enzyme mean

A

refers to the bacterial species from whcih the enzyme was isolated (Eco for E.coli)

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13
Q

what does the 4th letter of the restriction enzyme refer to

A

the strain of bacteria from which the enzyme was isolated

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14
Q

what doe sthe roman numerals that follow the letters mean

A

identify different enzymes from the same spe cies

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15
Q

restriction sites thend to be how long and generally

A

about 4-8 bp long and generally palindromic

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16
Q

the ends of restruction enzymes produced by cleavage are of which types

A

Cohesive ends ( 5’ overhangs or 3’ overhangs)
Blunt ends

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17
Q

overhangs are associated with which end

A

cohesive ends

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18
Q

even length ends from both single strands are which end

A

blunt end

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19
Q

the enzyme BamHi is from which microorganism
A. Clostridium formicoaceticum
b. Haemophilus aegyprius
c. Bacillus amyloliquefaciens

A

c. Bacillus amyloliquefaciens

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20
Q

how do restriction enzymes producing blunt ends and cohesive ends in DNA

A

by making double stranded cuts

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21
Q

cohesive ends are also known as

A

sticky. ends or overhanging ends

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22
Q

the nicks in the sugar phosphate backbone of two fragments can be sealed by

A

ligase

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23
Q

is it possible to insert DNA into a complex genome at single predetermined site using convention restriction enzymes?

A

NO

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24
Q

CRISP-CAS immunity is in

A

bacteria and archae

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25
Q

CRISP RNA are encoded by

A

Clustered Regularly Interspersed Short Palindromic Reoeats

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26
Q

A CSRISPR array is derived form

A

bacteriophage and plasmid genomes

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27
Q

______ combine with Cas protein to provide defense against invasion by specific foreign DNA molecules

A

CRSIPR RNAs

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28
Q

After expression and processing what happens to the foreign DNA

A

crRNA will join Cas to form an effector complex that binds to foreign DNA through base pairing

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29
Q

what makes the DNA nonfunctional after expression and processing

A

when the Cas protein cleaves to the foreign DNA

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30
Q

CRISPR-Cas 9 is a technique for ______

A

precisely editing the genome

31
Q

advantages of CRISP-Cas 9

A
  • can be used in intact cells
  • can be used in many species, including humans
  • relatively long sequence recog. by sg RNA (Edits can happen almost everywhere in genome
  • easier and less expensive than altering DNA binding protein
  • modified to introduce single cuts and sticky ends
32
Q

having single-stranded cuts and sticky ends through crisp-cas 9 means

A

improves efficiency of inserting DNA fragments

33
Q

Waht are some limitation and challenges of CRISPR-Cas 9

A

potential for off-target cleavage
genetic mosaic may result when applied to multicellular embryos
difficulty introducing the system into cells

34
Q

is Cas9 100% EFFICIENT

A

NO, some cells are edited while others are not

35
Q

Concerns with use of CRISPR-Cas 9 editing

A
  • could be modifying humans in questionable ways
  • already been used on human embryos but they did not complete development
  • germ line editing allows traits to be passed down through generations
  • Safety concerns because of off target cuts
  • potential danger of editing animals and plants that are released into the wild
36
Q

what method is used to seperate DNA molecules on the basis of their size

A

Gel electrophoresis

37
Q

after gel electrophoresis, DNA fragments appear as

A

bands on the gel

38
Q

which method can detect a single restriction fragment in a complex mixture of restriction fragment

A

Southern blotting

39
Q

in southern blotting, what DNA fragment will give signal on an autoradiogram

A

the fragments hybridized to a labelled probe

40
Q

what are the types of gel blots

A
  1. SOuthern blots
  2. Northern blots
  3. Western blot
41
Q

in which gel blot, is the DNA on the gel transferred to a filter then hybridized with a DNA or RNA probe

A

Southern blot

42
Q

in which gel blot, is the RNA on the gel transferred to a filter then hybridized with a DNA or RNA probe

A

northern blot

43
Q

in which gel blot is the protein on the gel transferred to a filter, an antibody is bused to detect a specific protein

A

Western blot

44
Q

specified DNA fragments can be amplified through

A

PCR (Polymerase Chain reaction),
molecular cloning

45
Q

what is an application of PRC

A

real time PCR

46
Q

the process of quantitavely detrmining the amount of DNA amplified as the reaction proceed is called

A

Real time PCR

47
Q

What are the limitation of PCR

A
  1. Sequence of the target DNA must be known2.
  2. Amplification of contaminants
  3. Accuracy
  4. Length of the PCR products
48
Q

in molecular cloning, how is a specific piece of DNA amplified

A

using a host cell

49
Q

what were used to replicate the desired piece of DNA in the host cell

A

Cloning vectors

50
Q

Examples of gene cloning

A

plasmid vectors
transformation
screening cells for recombinant plasmids
other cloning vectors
genetic engineering of plants for inset resistance

51
Q

circular DNA molecules from bacteria are called

A

Plasmids

52
Q

how is foreign DNA inserted into plasmid

A

using restriction enzymes and DNA ligase

53
Q

what are synthetic DNA fragments containing restriction sites

A

linkers

54
Q

what are used to confirm whether or not the cells have been transformed

A

Selectable markers

55
Q

an ideal cloning vector must have

A
  1. Origin of replication recognized in the host cell
  2. Selctable marker
  3. One or more unique restriction sites
56
Q

pUC19 is an axmple of

A

cloning vector

57
Q

what gene is used to screen for bacteria containing recombinant plasmids

A

LacZ gene

58
Q

original plasmid (nonrecombinant) produces which enzyme

A

Beta-galactosidase which cleaves to X-gal,

59
Q

what does the colonies of bacteria with a recombinant plasmid looks like

A

they remain white because recombinnat plasmid cannot synthesize Beta-galcatosidase

60
Q

what causes colonies to turn blue

A

beta-galctosidase produces by original plasmid which cleaves to X-gal

61
Q

examples of cloning vectors

A

phages
cosmids
Bacterial Artifical Chromosomes (BACs)
Yeast Artifical Chromosomes
Ti plasmids

62
Q

to ensure transcription and translation, a foreign gene may be inserted into

A

an expression vector like E-coli

63
Q

what contains operon sequences that allow inserted DNA to be translated and transcribed, and also sequences that regulate

A

expression vector

64
Q

gene-encoding repressor in expression vector does what

A

bind O (operator and regulates P(bacterial promoter sequences)

65
Q

What plasmid from Agrobacterium can be used to transfer genes into plants

A

Tiplasmid

66
Q

what is a gram-positive bacterium that produces several substances toxic to insects

A

Bt (Bacillus thuringiensis)

67
Q

Bt toxicity is specific to_______ and expressed in _____ to produce ______

A

insects, plants, insect resistant plants

68
Q

most notable insecticidal chemicals in Bt are

A

crystalline protein (cry called DElta-endotoxins)

69
Q

the Bt ttoxin gene was isolated from _____ and transferred to ____

A

bacteria, transferred to tobacco plants

70
Q

Transgenic tobaccos expressing Bt is protected from damage by ____

A

tobacco hornworm larvae

71
Q

DNA hybridization probes can be used to locate the

A

chromosomal or cellular location of a gene or its RNA product

72
Q

what is cDNA

A

DNA copy of mRNA

73
Q

how is cDNA made double stranded

A

DNA Polymerase

Genetics (20 decks)

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