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Recombinant DNA Flashcards

0
Q

Translation

A

Making proteins from information coded by DNA (via RNA intermediate)

RNA->Protein

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1
Q

Transcription

A

The process of synthesizing RNA by using one strand of a DNA molecule as a template.
DNA->RNA

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2
Q

Language of DNA and RNA

A

Nucleotides

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3
Q

Language of Protein

A

Amino acids

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4
Q

__ nucleotides specify __ amino acid

A

3, 1

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5
Q

Where does transcription happen in Eukaryotic Cell?

A

Nucleus

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6
Q

Where does Translation happen in Eukaryotic Cell?

A

The cytosol

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7
Q

Reasons for Cloning DNA?

A
  1. To study the DNA
  2. For DNA modification/mutation
  3. To transfer into cells/organisms
  4. For preservation/storage-freezing
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8
Q

Molecular Scissors

A

Restriction endonuclease (restriction enzymes)

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9
Q

Molecular Glue

A

DNA ligase

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10
Q

Where do restriction endonuclease come from?

A

Bacteria used for defense

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11
Q

Where is DNA ligase used?

A

Used in cells to put two pieces of DNA together

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12
Q

What does Enzyme produce?

A

A staggered cut that generates “sticky” or “cohesive” ends

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13
Q

EcoRI, Pstl, Smal, Hpall are examples of what?

A

Genus species enzymes

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14
Q

Making Recombinant DNA

A

Cut DNA with “molecular scissors” (restriction endonuclease)
Mix cut DNA’s together in test tube
Add “molecular glue” (DNA ligase) to form bonds between DNA’s

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15
Q

T or F: restriction endonuclease and DNA ligase are used to make recombinant DNA

A

T

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16
Q

Steps in DNA cloning

A
  1. DNA inserted into transfer vehicle (vector)
  2. DNA moved into host cell (transformation)
  3. Identical DNA copies made in host cell (DNA clones)
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17
Q

Cloning vectors are used to:

A
  1. Insert a DNA sequence of interest
  2. Stabilize a DNA of interest
  3. Move to a host cell to increase amount of DNA (cloning)
  4. Sometimes “express” the gene to make a protein and change characteristic
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18
Q

Characteristics of a Cloning Vector:

A
  1. Replicates (copied) in host cells
  2. Contains unique sites to insert DNA of interest (cloning sites)
  3. Features that allow the selection of cells that have the vector with the DNA of interest (selection genes)
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19
Q

Examples of Types of Cloning Vectors:

A
  1. Bacterial plasmids
  2. Viruses
  3. Artificial chromosomes (yeast, bacteria, human)
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20
Q

Artificial chromosomes

A

Laboratory constructs that perform functions of natural chromosomes

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21
Q

Artificial chromosomes applications

A
  1. To introduce DNA into a cell
  2. To study how chromosomes function
  3. Used in large DNA sequencing projects
  4. Future? Correct chromosomal defects
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22
Q

Proteins

A

Polymer of amino acids bonded together (covalent bonds)

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23
Q

Formation of strong bonds:

A

Subtract water (dehydration)

24
Breakage of strong bonds
Add water (hydrolysis)
25
RNA polymerase
Makes a polymer using the info from the gene. (Via RNA intermediate)
26
Viruses
Protein coat, sheath, core, cell wall Every form of life has a specific virus
27
Gene therapy
Create a genetic defect
28
Telomeres in chromosomes
People Age as Telomere shorten
29
Duplicated Chromosome
Duplicates it's cell chromosome before its divison
30
Somatic chromosomes
22
31
Chemical transformation Method
Animal and Plant Cells only Common, used to move DNA from animal cell to plant cell Shocks the cell (heat shock)
32
Electroporation transformation method
Electric pores flowing into membranes Reforms membrane so channels go through it. Transform bacteria cells, plant cells (w/ cell wall), animal cells
33
Gene Gun-Biolistics
Coats bbs with DNA Shoots bbs into cells, DNA goes in and come off bbs Can shoot directly in tissues, gene therapy while using gun
34
Liposome transformation method
Mediated transformation | Any cell that has membrane can use this to deliver goods
35
Micro injection transformation method
Use to inject eggs into animals Genetically modify animals Use micromanipulator Not for plants or fungi
36
Crown Gall Disease
When plants get cancer Immortal plant cells They grow and change forever Cause changes in plant cells
37
Ti plasmid
T-DNA is transferred to plant chromosomes | Main way to get transgenic plants
38
How do we know which cells have the new DNA?
Selection of host cells harboring recombinant DNA using specific methods Use of specific selection genes to accomplish this
39
Selection of Cells
Some genes used in selection are inactivated
40
How are some genes in selection inactivated?
By inserting foreign DNA into them. Gives cells with new DNA a specific characteristic that can be "visualized"
41
Intact vs. disrupted selection genes can be
Visualized
42
Antibiotic resistance genes
E.g. Ampicillin Resistance genes | Cells are antibiotic sensitive when gene inactivated
43
Genes that promote a cell color
Chromogenic
44
T or F: cells are colorless when gene is inactivated
T
45
Gene coding for an enzyme that breaks down compound in the medium
Turning cells blue
46
Complementary base pairing
Has to follow base pairing rules
47
mRNA
Coats for protein
48
Polymer(ASE)
Enzyme
49
Transfer RNA
Brings in amino acids to the growing protein chain
50
Central dogma
Start codon and stop codon
51
Ribosome
Bring in transfer RNA and has to match
52
Gene structure
RNA polymerase binds to promoter | When and where the gene turns on
53
2 ways that DNA ligase cut?
1. Sticky | 2. Cohesive
54
Making recombinant DNA
Have to have complimentary base pairing.
55
DNA ligase do?
Shielding cellular backbone together
56
Examples of vectors
Plasmids-transfer easily to other bacteria Viruses Ti-plasmids (plant vector)
57
Transformation
Move DNA into host cell

BIO 101 (3 decks)

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