Question
- Describe the characteristics a cloning vector must possess.
- Why DNA cannot pass through the cell membrane? Explain. How is a bacterial cell made ‘competent’ to take up recombinant DNA from the medium?
✓
Answer
- Should have ori/origin of replication,
- Has selectable marker, genes encoding for an antibiotic resistance/genes encoding for a - galactosidase,
- Has cloningsite/recognition site, for the restriction enzyme to recognise.
- DNA is a hydrophilic molecule
Bacterial cell is made competent by treating with specific concentration of Ca++ ion/divalent ions, incubating the mon ice, heat shock for a short period and placing it back on ice again.
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