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Molecular Biology > Exam 1 Content > Flashcards

Exam 1 Content Flashcards

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AP® Biology flashcards Biology 101 flashcards
1
Q

What is molecular biology?

A

The study of gene (and protein) structure and function.

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2
Q

Essential toolbox for molecular bio

A
  1. Cell culture
  2. PCR & cloning
  3. DNA sequencing & BLAST
  4. SDS-PAGE & Western Blot (immunostaining)
  5. Genome modification
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3
Q

Proteins are the _____ of western blot

A

Workhorse

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4
Q

What do the dark bands in a western blot represent?

A

A lot of protein is present

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5
Q

What does it mean when a band in a western blot go from darker to lighter (and vise versa) over time?

A

darker –> lighter = specific protein expression decreases

lighter –> darker = specific protein expression increases

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6
Q

What toolbox does western blot involve?

A

Cell culture, SDS-PAGE, and western blot

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7
Q

Cell culture….

A
  • Source material (human, transformed organism, unmodified organism)
  • Allows propagation of biomolecule of interest
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8
Q

PCR & Cloning…..

A
  • Making millions of copies of a DNA specific sequence

-Manipulating DNA sequence

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9
Q

DNA Sequencing…

A
  • Obtaining the sequence of monomers in a biomolecule
  • BLAST: search tool database, used to compare DNA sequences for similarities between organisms
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10
Q

SDS-PAGE & Western Blot…

A
  • Allows separation of molecules based on size
  • Allows visualization of a specific biomolecule in a mixed population of molecules
  • Antibodies used to find specific proteins
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11
Q

Immunostaining….

A
  • Using conjugated antibody to visualize a biomolecule of interest
  • EXAMPLES (Western Blot, ELISA, Immunohistochemistry, Immunocytochemistry)
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12
Q

ELISA…

A

Testing for molecules in a fluid (urine, blood, spit, etc.)

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13
Q

Immunohistochemistry….

A

Testing for molecules in a tissue

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14
Q

Immunocytochemistry….

A
  • Testing for molecules in a cell, and allows us to see whether they are in cytoplasm or nucleus
  • Toolbox: DNA manipulation, cloning, cell culture, immunocytochemistry
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15
Q

Stem cells have _____ that allow them to not differentiate into different cells

A

Masters regulators

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16
Q

Example: Western Blot –> Irreversible Post-Translational Modifications

A
  • Proteins that get cleaved
  • Tested by comparing band intensity of protein before its cleaved to band intensities of protein subunits after cleaved
17
Q

DNA synthesis in vitro

A

Arthur Kornberg

18
Q

RNA synthesis in vitro

A

Jerard Hurwitz

19
Q

Protein Synthesis in vitro

A

Paul Zemecnik

20
Q

Marshall Nirenberg

A

Cracked the genetic code

21
Q

DNA composition and structure (people)

A

Chargaff, Franklin, Watson, Crick

22
Q

Transformation (people who tested for it)

A

Griffith, Avery, Hershey and Chase

23
Q

DNA Replication (proved mechanism)

A

Meselson and Stahl

24
Q

PCR and DNA sequencing (people)

A

Mullis and Sanger

25
Q

Transforming Principle

A

A substance that can be transferred from nonliving cells to living cells, causing living cells to show characteristics of the nonliving cell.

26
Q

Frederick Griffith experiment (explain)

A

transformed a harmless strain of streptococcus (rough strain) into a pathogenic strain (smooth), but he didn’t show which biological molecule was responsible.

1.) Rough strain injected into mouse = mouse lives (rough strain recovered)

2.) Smooth strain injected into mouse = mouse died (smooth strain recovered)

3.) Smooth strain cell-free extract + rough strain injected into mouse = mouse died (smooth and rough strain recovered)

27
Q

Oswald Avery Experiment (explain)

A

Identified (indirectly) the “transforming principle” is a nucleic acid using enzymes

1.) Smooth strain cell-free extract + rough strain injected into mouse + DNAse = only rough strain recovered

2.) Smooth strain cell-free extract + rough strain injected into mouse + w/o DNAse = both rough and smooth strain recovered

28
Q

Stable Isotopes

A

Distinguished/identified only by a difference in mass

29
Q

Unstable isotopes

A

Identified by measuring radioactivity/energy that they release

30
Q

Centrifuge

A

Instrument that rotates samples at high velocities to separate components by denisty

Pellet = high density

Supernatant = lower density

31
Q

Alfred Hershey and Marsha Chase experiment (explain)

A

Identified (directly) the transforming principle as a nucleic acid using unstable isotopes and bacteriophages

1.) Generate two batches of radioactive phage. One with radioactive methionine (protein coat of bacteriophage) and the other with radioactive bacteriophage DNA

2.) Binding reaction (radioactive phage + bacterial cells)

3.) Knocked virus off bacteria used a blender

4.) Identify which isotope entered bacteria using a centrifuge
- Phage grown with radioactive protein coat = radioactivity in supernatant (not in cell)
- Phage grown with radioactive DNA = radioactivity in pellet (inside cell)

32
Q

Matthew Meselson and Franklin Stahl experiment (explain)

A

Showed that DNA replication was semiconservative using stable isotopes

1.) Grow bacteria in media containing N15 (heavy isotope) = lots of DNA containing N15 after replication

2.) Shift bacteria to media containing N14 (light isotope) = DNA shifting from containing N15 only to containing both N15 & N14, then after time containing majority N14 and little N15

33
Q

What are cell-free extracts?

A

Content inside a cell that is needed to replicate DNA or express a gene

34
Q

Kary Mullis experiment (explain)

A

Used a thermostable DNA polymerase to make PCR more efficient

High temps are required to denature DNA strands. A thermostable bacteria contains a DNA polymerase that is not denatured at high temps and can function at high temps.

35
Q

Modified Nucleotides

A

Chain terminating nucleotides that lack the 3’-OH group that is required for the formation of the phosphodiester bond between two nucleotides.

This causes DNA polymerase to cease synthesis of the new DNA strand.

Fluorescently or radioactively labeled for detection

36
Q

Fredrick Sanger experiment (explain)

A

Used chain terminating nucleotides to read the sequence of a DNA molecule

1.) 4 separate sequencing rxn’s, each w/ one of 4 fluorescently conjugated chain terminating nucleotides

2.) 4 groups of fragments that end in 1 of 4 colors (bases). “Read” the DNA sequence from the bottom of the gel to the top of the gel.

37
Q

DNA Replication vs. PCR

A

DNA replication:
- in vivo
- Duplicate entire genome (2x)
- involve enzymes & accessory proteins
- use helicase for DNA strand separation

PCR
- in vitro
- amplify a specific DNA sequence (2x)
- involves some enzymes
- use high heat for DNA strand separation

Molecular Biology (3 decks)

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