Problems with AU1tag

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Thu Feb 8 09:07:33 EST 2001

This message has been posted by:  "Luis M. Martins" <L.Martins at>

Hi I have tried to insert an AU1 tag either at the N- or C-term of my
protein by PCR. I have inserted the sequence met-DTYRYI-GG-ORF or
ORF-GG-DTYRYI-end into pcDNA3. I have sequenced and things seem OK. I tried
to check for transient expression in 293 cells.

I was very surprised not to detect any protein by western using an anti-AU1
Ab. My control with a previously AU1-tagged protein worked (this construct
wasn't made by me)

Any thoughts?

Thank you


Luis Miguel Martins, Ph.D.
Signal Transduction Laboratory
Imperial Cancer Research Fund
44 Lincoln's Inn Fields
London WC2A 3PX

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