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How are restriction enzymes (endonucleases) isolated from all other periplasmic proteins?

julesarth

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Posts: 105

Rep:

11 years ago

How are restriction enzymes (endonucleases) isolated from all other periplasmic proteins?

When a gram-negative cell is osmotically shocked to release it's perplasmic contents, how are restriction endonucleases isolated?

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riveraj22

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#1

11 years ago

I don't know how much you know about lab protocols for different things, but you can use ion exchange chromatography. Just google it if you don't know how it works. Column fractions are assayed for the enzyme using Lambda DNA and digestion products are identified by agarose gel electrophoresis. Fractions that contain the wanted restriction enzyme are identified and pooled.

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