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Module 6.1 · Topic 9
Module 6.1 · Topic 9
Moving a gene between vectors without redesigning cut sites every time
A gene successfully cloned into a bacterial expression vector now needs testing in a mammalian system too — a different vector entirely. Traditional restriction-and-ligation cloning means finding new compatible cut sites and, quite possibly, discovering the gene's own internal sequence conflicts with the enzyme chosen. Every new vector risks starting over.
Instead of restriction sites, a cloning system uses standardised recombination sequences flanking a gene of interest, paired with matching sequences on many different destination vectors. Could this let one entry clone move into multiple different vectors using the same simple reaction each time?
Recombination-based cloning systems, such as Gateway cloning, offer an advantage over traditional restriction-ligation cloning by:
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