Introduce ribosomal binding site (RBS) in pET?? - (Jun/29/2005 )

How to introduce ribosomal binding site (RBS) sequence together with my gene into pET plasmid by PCR?? How to design the primer??

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Hi friend,
You can do it by simply add in the RBS sequence into your sense primer sequence.
Let say:
AGGAGGAACAGCAAUGNNNNNNNN

where the underlined sequence is RBS and bolded sequence is initiator sequence, followed by NNNNNN which is your primer.

Note:
The sequence above is Shine-Dalgarno sequence (prokaryotic RBS).

Regards
Hadrian

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