fusion protein expression

yunkiam k-yap at bs.aist-nara.ac.jp
Wed Apr 5 23:00:20 EST 2000


Dear netters,

I am a PhD student in Japan.  I am facing some problems in expressing my
fusion proteins (both GST-fusions and His-tag fusions) in soluble
fractions; therefore I decided to use  the TNT-coupled recticulocyte
system to synthesis the desired proteins.

I need to use these fusion proteins for the following assay:

(i) protein interaction - pull down assay
(ii) in-vitro kinase assay

My questions are:

(i) how to purify the translated product from the TNT- coupled
recticulocyte system?
(ii) Are there any references for the above assays (pull down assay &
in-vitro kinase assay) using both in-vitro translated fusion proteins?

Your help are kindly appreciated. Thank you.

sincerely,
YK

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