Lab 5 Flashcards

1
Q

Where do restriction enzymes cut?

A

At the DNA backbone.

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2
Q

Restriction enzyme site:

A

The target site from which the DNA of interest is attached to the plasmid.

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3
Q

Plasmid:

A

Extra-chromosomal DNA that replicates independently of a chromosome.

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4
Q

A recombinant plasmid:

A

A plasmid that contains genes from two different sources.

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5
Q

DNA ligase:

A

Glues two pieces of DNA together.

It uses Phosphodiester bonds to connect the backbones together and ATP is used as an energy source.

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6
Q

Another name for a plasmid:

A

Vector.

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7
Q

What does a vector not have:

A

An insert.

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8
Q

Most plasmids contain:

A

Antibiotic resistance gene.

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9
Q

Plasmids’ ORI:

A

Origin of replication.
This is needed for the plasmid to be replicated in the bacterium.
ORI is where replication begins.

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10
Q

Polylinker / Polycloning:

A

Contains several restriction enzyme sites that are very close together.

This is where the gene is inserted.

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11
Q

Bacteria Transformation:

A

The process of moving a plasmid into a bacteria.

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12
Q

pHUG21 contains:

A

A heme transport gene.
An ampicillin resistance gene.

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13
Q

Another name for restriction enzyme:

A

Restriction endonuclease.

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14
Q

Restriction endonuclease:

A

An enzyme that cuts the DNA at a specific DNA sequence within the DNA strands, rather than at the very end.

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15
Q

Types of ends a restriction enzyme can leave:

A

Sticky end [Cohesive/overhang]
Blunt end.

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16
Q

Another name for sticky end:

A

Cohesive end.
Overhang.

17
Q

Palindromic sequence:

A

A sequence that reads the same backward and forwards.

18
Q

Restriction enzymes cut at:

A

Specific sites.

19
Q

Restriction enzymes recognize:

A

The entire sequence.

20
Q

Blunt end:

A

No overhang, harder to hydrogen bond.

21
Q

How many more pieces of inserts than plasmids do you need?

A

4 times as many.

22
Q

Why do bacteria make restriction endonuclease:

A

Bacteria produce them to chop up DNA from a bacteriophage that may have infected the bacterium.

23
Q

The plasmid and the DNA need to have the same:

A

Overhangs.

24
Q

Methylase:

A

Methylate some of the bases in the sequence so the restriction enzyme cannot cut it.

25
Restriction enzyme modification:
Made up of the restriction enzyme and the methylase.
26
Why is modification done:
To prevent degradation of the host DNA by its restriction endonuclease.
27
Where is methylation done:
On the bases.
28
Restriction endonuclease nomenclature Ex: E. coli RY13 1.) The first letter represents: 2.) The 2nd and 3rd letters represents: 3.) The second capital letter represents: 4.) The number represents:
EcoR1 1.) The genus. 2.) Species the enzyme was isolated. 3.) The strain. 4.) The order in which the enzyme was discovered.
29
How do molecular biologists use restriction enzymes:
They use them to cut up DNA of interest into smaller pieces which are easier to manipulate.
30
Restriction enzyme cuts:
On both sides of the gene of interest.
31
kB stands for:
Kilobases.
32
Recombinant plasmids can be used by researchers to:
Overexpress a gene so protein can be purified and studied more closely.
33
Expression vectors:
Vectors that are used to overexpress a gene.
34
Restriction enzyme map can:
Help you create a plasmid that contains only a certain gene.