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Biology reviewer > RECOMBINANT DNA TECHNOLOGY > Flashcards

RECOMBINANT DNA TECHNOLOGY Flashcards

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AP® Biology flashcards Biology 101 flashcards
1
Q

TECHNOLOGY THAT ENCOMPASSES A NUMBER O F EXPERIMENTAL PROTOCOLS
LEADING T O TRANSFER O F DNA FROM ONE ORGANISM T O ANOTHER

A

Recombinant DNA TECHNOLOGY

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2
Q

Molecular scissors that cut DNA into fragments at specific sites in their sequence.

A

Restriction endonucleases

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3
Q

EcoRI
“Eco” word from?

A

1st letter of genus +
1st and 2nd letter
of species

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4
Q

EcoRI
“R”from ?

A

Strain of host bacteria

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5
Q

EcoRI
“I”from ?

A

Order of discovery

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6
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A
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6
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7
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9
Q
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10
Q

CLASSIFICATION OF RESTRICTION ENZYMES

Cut DNA at random far from their recognition sequences.

A

Type 1

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10
Q

Cut DNA at defined positions close to or within their recognition sequences.

A

Type 2

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10
Q

Cut modified (e.g. methylated) DNA.

A

Type 4

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11
Q

Cut DNA outside of two recognition sequences in opposite orientations.

A

Type 3

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12
Q

MECHANISM OF RESTRICTION ENZYMES
1.
2.
3.

A
  1. SCAN - Look fo a particular pattern of
    nucleotides:ENZYME’S RECOGNITION
    SEQUENCE
  2. BIND- The enzyme will bind the DNA
    molecule.
  3. CUT- The enzyme cuts in each of the two
    sugarphosphate backbones of the
    double helix
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13
Q

Alul and Haelll produce _____ ends.

A

blunt

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14
Q

DNA sequence that produce blunt ends.

A

Alul and Haelll

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15
Q

BamHl, Hindlll and EcoRi produce ____ ends

A

sticky

16
Q

Made by fusing DNA-binding domain to a nuclease domain.
Ex. CRISPR technolog

A

Artificial Restriction enzymes

16
Q

DNA sequence that produce “ sticky” ends

A

BamHl, Hindlll, and EcoRi

17
Q

Who won nobel prize in Chem 2020, “for the development of a method for genome editing.

A

Emmanuelle Charpentier
Jennifer A. Doudna

18
Q

A DNA MOLECULE IN WHICH FOREIGN DNA
MOLECULE IS INSERTED AND WHICH IS FURTHER CAPABLE OF REPLICATING WITHIN HOST CELL TO PRODUCE MULTIPLE CLONES OF THE RECOMBINANT DNA.

A

CLONING VECTOR

19
Q

MAIN FEATURES OF CLONING VECTORS

A
  1. ORIGIN OF REPLICATION
  2. MULTIPLE CLONING SITES
  3. SELECTABLE MARKER
20
Q

DNA segment with several RE sites located next to each other.

A

MULTIPLE CLONING SITE

21
Q

Allows selection of host cells with the recombinant vector.

A

Selectable marker

22
Q

CLONING STRATEGIES

_________ involves the ligation of short, staggered ends that can be rejoined by DNA ligase to regenerate original recognition
sequence.

A

Cohesive End Cloning

23
Q

CLONING STRATEGIES

_____ involves ligating dsDNA
into a plasmid where both the insert and
linearized plasmid have no overhanging bases
at their termini.

A

Blunt End Cloning

24
Q

10-100x less efficient than cohesive-end
cloning. Desired insert does not require any restriction sites.

A

Blunt End Cloning

25
Q

CLONING STRATEGIES

Modified Blunt End Cloning may use…

A
  • Linkers
  • Adaptors
  • Homopolymer tails
26
Q

final step in the construction of a recombinant
DNA molecule where the vector molecule and
the DNA to be cloned are joined together by
DNA ligase.

A

LIGATION

27
Q

Two families of dna ligases

A

1.ATP-dependent Ligase
2. NAD+-dependent ligases (LigA)

28
Q

All known eukaryal cellular DNA ligases; also in archaea.

A

ATP-dependent Ligase

29
Q

distinctive and structurally homogeneous clade of enzymes found in all bacteria.

A

NAD+-dependent ligases (LigA)

30
Q

is the process by which an organism
(competent) acquires exogenous DNA.

A

TRANSFORMATION

31
Q

It is a Direct Methods of transformation,

  • CaCl2
  • TSS Buffer with PEG, DMSO, and MgCl2
  • Usually followed by heat-shock transformation.
A

Chemical Competency

32
Q

It is a Direct Methods of transformation,

_____ significantly increases the electrical
conductivity of cell membrane for
subsequent increase in its permeability.

A

Electrocompetency

33
Q

It is a Direct Methods of transformation,

the use of a glass micropipette to inject a liquid
substance at a microscopic or borderline
macroscopic level.

A

Microinjection

34
Q

It is a Direct Methods of transformation,

  • Transforming yeast cells by limited physical
    disruption in the presence of DNA
  • This method is less efficient but considerably
    more convenient
A

Agitation by glass beads

35
Q

4 INDirect Methods of transformation.

A

Transduction
Agrobacterium Mediated
Protoplast Fusion
Conjugation

36
Q

2 METHODS OF SELECTION

A

1.Use of markers
2.Use of reporter genes

37
Q

ANTIBIOTICS
-Positive selection
* Ex. Ampicillin

under what methods if selection?

A

METHODS OF SELECTION: 1. USE OF MARKERS

38
Q

Negative selection

A

BLUE-WHITE SCREENING

39
Q

The genes encode protein products that can
be observed using certain biochemical
assays.

A

METHODS OF SELECTION: 2. Reporter genes

40
Q

Examples of Reporter genes. The genes encode protein products that can
be observed using certain biochemical
assays.

A

– β-glucuronidase (GUS)
– Luciferase gene
– Green Fluorescence Protein (GFP)

Biology reviewer (6 decks)

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