Unit 1: Cell And Cell Research Flashcards

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1
Q

Define a cell.

A

The cell is the basic structural, functional, and biological unit of all known organisms. The cell is the minimum unit of an capable of organism acting autonomously

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2
Q

What are the 4 rules of cell theory.

A
  1. All living organisms are composed of one or more cells.
  2. The cell is the basic unit of structure and organisation in organisms.
  3. Cells arise from pre-existing cells.
  4. Hereditary information (DNA) is passed on from cell to cell.
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3
Q

When was cell theory formulated?

A

1839

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4
Q

Who invented the first microscope and what did they see?

A

Antonie van Leeuwenhoek saw some cells (Protozoa, blood cells, sperm cells and bacteria)

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5
Q

When was Antonie van Leeuwenhoek alive?

A

1632-1723

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6
Q

Define prokaryotic cells.

A

Derived from the Greek “before the nucleus”. They comprise bacteria and archaebacteria and are small cells with simple structure. Absence of a nucleus so the genomes are less complex and do not contain cytoplasmic organelles or cytoskeleton.

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7
Q

Define eukaryotic cells.

A

Derived from the Greek “true nucleus” and make up all other living organisms including protozoa, fungi and animals. They are much bigger.

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8
Q

Spontaneous formation of organic molecules experiment.

A

Water vapour was expelled into an atmosphere made up of hydrogen (H2), methane (CH4) and ammonia (NH3) in which sparks were discharged. Analysis of the reaction products revealed the formation of several organic molecules.

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9
Q

Name some of the products.

A

Alanine, aspartic acid, glutamic acid and glycine.

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10
Q

Who carried out/discovered this experiment?

A

Stanley Miller

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11
Q

Formation of macromolecules

A

Monomers have been shown to spontaneously polymerise under plausible prebiotic conditions. E.g heating dry mixtures of amino acids results in polypeptides.

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12
Q

What is the fundamental characteristic of a macromolecule for life to evolve?

A

The ability to replace itself AKA the synthesis of new copies of itself.
**Only nucleic acids (DNA and RNA) are capable of directing their replication

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13
Q

RNA self replication

A

Complementary pairs between nucleotides ( adenine with uracil and guanine with cytosine) allow an RNA strand to serve as a template for the synthesis of a new strand with the complementary sequence. RNA is the only one capable of catalysing its own replication.

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14
Q

What is the central dogma of molecular biology?

A

How proteins are formed.

  • DNA to RNA = transcription
  • RNA to protein = translation
  • RNA to DNA = reverse transcription (tRNA read rna to dna?)
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15
Q

How is the first cell believed to have arisen?

A

From the coating of self-replicating RNA and its associated molecules by a membrane composed of phospholipids.

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16
Q

What would the first cell have consisted of?

A

Self-replicating RNA and it’s encoded proteins.

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17
Q

What does each phospholipid molecule contain?

A

2 long hydrophobic tails attached to a hydrophilic head. The hydrophobic tails are embedded in the lipid bilayer; the hydrophilic heads are exposed to water on both sides of the membrane.

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18
Q

Generation of metabolic energy

A
  • Glycolysis: glucose (C6H12O6)-> lactic acid (2 C3H6O3) and generates 2 ATP.
  • Photosynthesis: 6 CO2 + 6 H2O -> C6H12O6 + 6 O2.
  • Oxidative metabolism: C6H12O6 + 6 O2 -> 6 CO2 + 6 H2O and generates 36-38 ATP.
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19
Q

What is glycolysis? (evolution of metabolism)

A

The anaerobic breakdown of glucose into lactic acid,.

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20
Q

What is photosynthesis? (evolution of metabolism)

A

Uses the energy from the sun to synthesise glucose from carbon dioxide and water, releasing oxygen ans a product. This O2 is used by oxidative metabolism.

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21
Q

What is oxidative metabolism? (evolution of metabolism)

A

The breakdown of glucose into carbon dioxide and water, releasing more energy than from glycolysis.

22
Q

What is Escherichia coli?

A

A rod shaped bacterium that usually inhabits the intestines of humans and other vertebrates

23
Q

What is in the rapid growth medium for E.coli?

A

Glucose, salts and other organic compounds e.g. amino acids, vitamins, nucleic acid precursors

24
Q

What is in the slow growing medium for E.coli

A

Salts, a nitrogen source (e.g. ammonia) and a carbon and energy source (e.g. glucose)

25
Q

What is fission?

A

Symmetric form of asexual reproduction: consists of duplication of DNA, then the division of cytoplasm (cytokinesis), resulting in 2 daughter cells

26
Q

What is budding?

A

Asymmetric form of asexual reproduction: a protrusion is created in a cell and continues to grow, leading to another cell that is (at first) smaller than the initial cell

27
Q

What is humanised yeast?

A

When human genes are introduced into yeast or it’s own genes are replaced by human orthogonal to study their functioning

28
Q

What can humanised yeast be used for?

A

The pharmaceutical industry use them to design new drugs that interact with the product of expressed human genes (an inhibitor of an enzyme or a ligand for a receptor). Analysis of physiological effects on the microorganism aids early stages of drug development

29
Q

What won the Nobel prize in physiology in 2001?

A

The discoveries of key regulators of the cell cycle, found using a yeast model, (by Leland H. Hartwell, Tim Hunt and Sir Paul M. Nurse)

30
Q

What won the Nobel prize in physiology in 2013?

A

Vesicle trafficking (by Randy W. Schekman) - for their discoveries of machinery regulating vesicle traffic, a major transport system in our cells

31
Q

What has Caenorhabditis elegans (nematode) been used to study?

A

The genetics of animal development and cell differentiation and the nervous system

32
Q

What is Caenorhabditis elegans being used for recently?

A

It contributes to the causes of aging, cell death and the structure of the genome

33
Q

Which model organism has won 3 Nobel prizes?

A

C. elegans for apoptosis by Brenner, Sulston and Horvitz

34
Q

Which universal genetic tools were pioneered and by which model organism?

A

C. elegans helped develop:

  • RNAi (RNA interference) - (mechanism of regulation of genes that used as a technique for the functional analysis of the genome allows to introduce well-defined sequences of double stranded RNA into worm to silence almost any gene
  • Green Fluorescent Protein (GFP) - transparency of C. elegans allowed use for in vivo monitoring of certain biological processes
35
Q

What is magnification?

A

The factor by which an image appears to be enlarged

36
Q

What is resolving power?

A

The ability of the objective lens to distinguish as separate and different two points very close to each other

37
Q

What is the maximum magnification of a light microscope and electron microscope?

A
  • light microscope: X1000

- electron microscope: X150,000 - X200,000

38
Q

What is the resolving power of the human eye, a light microscope and a transmission electron microscope?

A
  • Human eye: 0.1 mm
  • Light microscope: 0.2 μm
  • Transmission electron microscope: 1-2 nm (10-20 ángstrom (Å))
39
Q

What three systems make up a conventional optical microscope?

A
  • Mechanical: foot (base), column(arm), stage(sample holder) and tube (head)
  • Optical: objectives and eyepieces
  • Lighting: spotlight, condenser and diaphragm
40
Q

Magnification and resolution of optical microscope?

A
  • Magnification: about 1000 times (most cells can be seen)

- Resolution: 0.2 μm (does not allow to observe small details of cellular structure)

41
Q

What will give a clearer image?

A

A smaller distance (n)

  • n = refractive index of the medium e.g 1.0 for air + 1.4 for immersion oil
42
Q

Light field microscope

A
  • Ability to distinguish cell parts depends on contrast from absorption of visible light by cellular components
  • *Fixation: stabilise and preserve cell structures (alcohol, acetic acid, formaldehyde…)
  • *Staining: highlights contrast
  • Limitation: can’t use on live cells
43
Q

Phase contrast microscope

A
  • Can view live samples without staining
  • Manipulates light to increase contrast of the sample => can view structures that are “invisible: through conventional microscopes
  • Works by increasing the contrast of the phases of the waves reaching the objective (phase= measurement of the wave’s position relative to a reference point)
44
Q

How does the phase contrast microscope work?

A
  • Lighting method that treats a portion of the light (the one that undergoes diffraction through the object) differently to the rest (the portion that undergoes no alteration). This causes the first portion of light to interfere with the rest, so a visible image of a transparent sample is created.
  • Goal is to manipulate the phase shift of 2 light beams to translate these small phase changes into corresponding amplitude changes that are visualise as contrast differences
  • ** Small phase differences turn into intensity changes
45
Q

Which microscope is similar to the phase contrast microscope and why?

A

The Differential Interference Contrast (DIC) as it also manipulates light to create a contrast

46
Q

Differential Interference Contrast (DIC)

A
  • Gives pseudo-3D shaded images
  • Can be used on live, unstained samples
  • Highly sensitive to thin cellular material even when located in a thick tissue
  • Limitation: 3D images can be misleading in some cases e.g. could show bumps that look like structures within a cell that does not contain any and is actually an optical impression
47
Q

How does a DIC microscope work?

A
  • **Uses polarised light and prisms to convert phase delays into intensity changes (contrast)
  • Two waves are spatially shifted int he plane of the sample and recombined before observation
  • **The interference of two waves in recombination is sensitive to the optical difference in their optical path
48
Q

Fluorescence Microscopy

A
  • *Based on property of fluorescent substances (fluorochromes) to absorb light from a certain λ, and emit light from another λ
  • Used to see selectively labelled structures (with fluorescent substances ) + study of intracellular distribution of molecules in living of fixed cells
    ~ *Nucleic acids: Specific genes or RNA transcripts can be detected by hybridisation with complementary probes labelled with fluorescent marker
    ~ *Proteins: Can be detected with labelled antibodies that recognise the specific proteins to be studied, or by fusing with fluorescent proteins e.g GFP (green fluorescent protein)
49
Q

Widefield Fluorescence Microscopy

A
  • Light passes through *excitation filter to select the light from a λ that excites dye
  • Then a *dichroic mirror deflects the excited light to the sample
  • Light emitted by the sample passes through the mirror and a *barrier filter that selects the λ emitted by the fluorescent dye
50
Q

Green Fluorescent Protein (GFP)

A
  • Isolated from jellyfish
  • Can be fused to any protein of interest using recombinant DNA technology
  • Does not require fixation or staining, unlike use of antibodies
  • Allows proteins to be visualised in living cells
51
Q

FRAP: fluorescence recovery after photo bleaching

A
  • Method follows movement of GFP labelled proteins
  • Region of interest marked by GFP protein is bleached by high intensity light. Fluorescence recovers over time through movement of unbleached GFP labelled molecules into bleached region
  • Rate of fluorescence recovery provides rate of protein movement within cell