Genetic Cooking - Vectors

13 important questions on Genetic Cooking - Vectors

What are cloning vectors?

DNA molecules used to “transport” cloned gene.

What 3 properties would an ideal cloning vehicle have?

  1. Low molecular weight.
  2. Ability to confer readily selectable phenotypic traits on host cells.
  3. Single sites for a large number of restriction endonucleases.

What are the types of cloning vectors?

  • Plasmid
  • Phage
  • Cosmid
  • BAC, YAC
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What is the average size of plasmid vectors?

1,2 - 6 Kb.

What 3 things do all plasmid vectors contain?

  • Replication Origin (ORI) sequence.
  • A gene that permits selection.
  • Multiple Cloning Site (MCS): for insertion of exogenous DNA.

What is the structure of a phage vector?

Head, tail, tail fibers.

What is a phage vector?

A bacteriophage that inserts linear DNA with sticky ends in a host cell (E. coli). Here the sticky ends hybridize and the DNA becomes circular.

What is a cosmid vector?

A combination of the plasmid vector and the COS site which allows the target DNA to be inserted into the λ head. It can clone large inserts of DNA (45 Kb).

What is a BAC?

A bacterial artificial chromosome. It can be cloned as a plasmid in a bacterial host. It can clone up to a few hundred Kb of DNA.

What is a YAC?

A yeast artificial chromosome. It can take up to 1000 Kb of DNA. It has to be linear. The YAC can use both yeast and bacteria as a host.

What is a shuttle vector?

A vector that can replicate in both prokaryotic and eukaryotic cells.

What is the plasmid copy number?

The number of plasmids that can be inside of one cell.

What is plasmid incompatibility?

Incompatibility refers to the fact that different plasmids are sometimes unable to coexist in the same cell. This occurs if the two different plasmids share functions required for replication and/or partitioning into daughter cells. Direct competition for these functions often leads to loss of one of the plasmids from the cell during growth of a culture.

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