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Virology > 3+4: Nucleic Acids, Lipids, Carbohydrates > Flashcards

3+4: Nucleic Acids, Lipids, Carbohydrates Flashcards

1
Q

Characteristics of Viral Nucleic Acids:

A
  • Carries the genetic information, determines viral properties
  • Relatively small (3–300kb)
  • dsDNA, ssDNA, dsRNA, ssRNA
  • linear or circular
  • continuous or segmented
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2
Q

virions may contain alien nucleid acids like:

A
  • Polyomaviruses, pestiviruses: host cell nucleic acid
  • Arenaviruses: ribosomes
  • Retroviruses: onc, src genes
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3
Q

Investigation of RNA infectivity can be done by?

A
  • positive sense

- negative sense

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4
Q

What is positive sense?

A

5´to 3´ direction. Happens fast. It goes straight to mRNA, without the transcription process
- i. e: Picornavirus

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5
Q

What is negative sense?

A

3´ to 5´ direction. Slow. Need viral enzymes for transcription

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6
Q

What are the different types of electrophoresis?

A
  • Agarose-gel electrophoresis

- sometimes polyacrylamide-gel electrophoresis

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7
Q

Restriction endonuclease analysis consist of:

A
  • dsDNA

- Endonucleases

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8
Q

What are endonucleases?

A
  • Bacterial defense enzymes
  • recognize certain sequences
  • sticky end - blunt end
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9
Q

What happens in Restriction endonuclease analysis?

A
  • Enzyme cleavage

- Physical mapping

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10
Q

Characteristics of Enzyme cleavage:

A
  • more exact size
  • smaller fragments - handling
  • -> restriction fragment length polymorph
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11
Q

What are Restriction fragment length polymorph. used for?

A
  • Identification and taxonomy

- Epidemiological investigations

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12
Q

What are the function of Physical mapping?

A
  • Localization of cleavage sites

- partial digestion with endonucleases

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13
Q

Molecular cloning of viral DNA consists of?

A
  • Propagation of virus DNA fragments in bacterial plasmids
  • Mass DNA production: quicker, cheaper
  • Expression plasmids: protein production
  • safe bacteria are needed.
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14
Q

What is the role of Viral Proteins?

A
  • defense and targeting of the genome
  • shape of the virion
  • enzymes for multiplication
  • receptors
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15
Q

Grouping of proteins:

A
  • Structural proteins: surface proteins, core proteins.

- Non-structural proteins

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16
Q

What are/are the function of surface proteins?

A

Structural proteins

  • capsid, envelope
  • Shape, antigenicity, adsorption, HA activity, enzymes for penetration or or release of virions
17
Q

What are/are the function of Core proteins?

A
  • Na protection, Stabilization, enzymes for replication
18
Q

Where can you find the Non-structural proteins?

A
  • coded in the viral genome
  • not integrated in the virion
  • present only in the vegetative virus
19
Q

Types of Non-structural proteins:

A
  • Early, immediate-early proteins

- Late proteins

20
Q

What are/are the function Early, immediate-early proteins?

A

Non-structural proteins

  • regulation of the cell, inhibition of cellular defence
  • enzymes for replication
21
Q

What are/are the function Late proteins?

A

Non-structural proteins

  • enzymes for structural protein maturation, virus assembly
  • (the majority of the late proteins are structural)
22
Q

Name the methods of protein investigation:

A
  • SDS-Polyacrylamide-gel electrophoresis (PAGE)
  • Immunobloting (Western blot)
  • Immunoperoxidase staining
  • Monoclonal antibody production
23
Q

Characteristics of SDS-Polyacrylamide-gel electrophoresis (PAGE):

A
  • concentrated virus suspension
    • ## SDS + Mercapto-ethanol = Linearization (polypeptides)
24
Q

Staining used in SDS-Polyacrylamide-gel electrophoresis (PAGE)?

A

Silver, Coomassie Brilliant Blue dye.

  • Polypeptide map
  • characteristic to the virus
  • -> number and size of viral polypeptides
25
Q

Function of Immunobloting (western blot)?

A

Transfer of viral proteins from PAGE into nitrocellulose filter

26
Q

When do we use Immunoperoxidase staining?

A
  • SDS-PAGE
  • Western blot
  • histology sections
27
Q

Function of Immunoperoxidase staining?

A
  • Detection of antigens
  • Diagnostics, identification
  • mechanism and timing of protein production
  • epitope investigation
28
Q

Characteristics for Monoclonal antibody production?

A
  • MAb produced by a single clone of B cells
  • specific for a particular epitope
  • Cloning
  • Use for diagnostics and virus analysis
29
Q

What happens if you mix MAb with myeloma cells?

A

Mitosis, but no AB production

30
Q

Testing of the clones:

Monoclonal antibody production

A

ELISA, IF, IPO

–> epitope specific MAbs

31
Q

What are viral lipids?

A
  • enveloped viruses

- acquired from cellular membrane strictures (budding)

32
Q

What is the cellular membrane structures of the Viral lipids?

A
  • phospholipid, cholesterol
    • virus specific proteins, glycoproteins
  • -> poxviruses, ASFV
33
Q

What are viral carbohydrates?

A
  • ribose, deoxyribose, in the NA
  • Glycoproteins, on the surface
    pox: in the core to

Virology (25 decks)

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