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BIO 101 > Recombinant DNA > Flashcards

Recombinant DNA Flashcards

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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
Q

Breakage of strong bonds

A

Add water (hydrolysis)

25
Q

RNA polymerase

A

Makes a polymer using the info from the gene. (Via RNA intermediate)

26
Q

Viruses

A

Protein coat, sheath, core, cell wall

Every form of life has a specific virus

27
Q

Gene therapy

A

Create a genetic defect

28
Q

Telomeres in chromosomes

A

People Age as Telomere shorten

29
Q

Duplicated Chromosome

A

Duplicates it’s cell chromosome before its divison

30
Q

Somatic chromosomes

A

22

31
Q

Chemical transformation Method

A

Animal and Plant Cells only

Common, used to move DNA from animal cell to plant cell

Shocks the cell (heat shock)

32
Q

Electroporation transformation method

A

Electric pores flowing into membranes

Reforms membrane so channels go through it.

Transform bacteria cells, plant cells (w/ cell wall), animal cells

33
Q

Gene Gun-Biolistics

A

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
Q

Liposome transformation method

A

Mediated transformation

Any cell that has membrane can use this to deliver goods

35
Q

Micro injection transformation method

A

Use to inject eggs into animals
Genetically modify animals
Use micromanipulator
Not for plants or fungi

36
Q

Crown Gall Disease

A

When plants get cancer
Immortal plant cells
They grow and change forever
Cause changes in plant cells

37
Q

Ti plasmid

A

T-DNA is transferred to plant chromosomes

Main way to get transgenic plants

38
Q

How do we know which cells have the new DNA?

A

Selection of host cells harboring recombinant DNA using specific methods

Use of specific selection genes to accomplish this

39
Q

Selection of Cells

A

Some genes used in selection are inactivated

40
Q

How are some genes in selection inactivated?

A

By inserting foreign DNA into them. Gives cells with new DNA a specific characteristic that can be “visualized”

41
Q

Intact vs. disrupted selection genes can be

A

Visualized

42
Q

Antibiotic resistance genes

A

E.g. Ampicillin Resistance genes

Cells are antibiotic sensitive when gene inactivated

43
Q

Genes that promote a cell color

A

Chromogenic

44
Q

T or F: cells are colorless when gene is inactivated

A

T

45
Q

Gene coding for an enzyme that breaks down compound in the medium

A

Turning cells blue

46
Q

Complementary base pairing

A

Has to follow base pairing rules

47
Q

mRNA

A

Coats for protein

48
Q

Polymer(ASE)

A

Enzyme

49
Q

Transfer RNA

A

Brings in amino acids to the growing protein chain

50
Q

Central dogma

A

Start codon and stop codon

51
Q

Ribosome

A

Bring in transfer RNA and has to match

52
Q

Gene structure

A

RNA polymerase binds to promoter

When and where the gene turns on

53
Q

2 ways that DNA ligase cut?

A
  1. Sticky

2. Cohesive

54
Q

Making recombinant DNA

A

Have to have complimentary base pairing.

55
Q

DNA ligase do?

A

Shielding cellular backbone together

56
Q

Examples of vectors

A

Plasmids-transfer easily to other bacteria

Viruses

Ti-plasmids (plant vector)

57
Q

Transformation

A

Move DNA into host cell

BIO 101 (3 decks)

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