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[Protein-analysis] zinc finger protein purification issue

김종문 via proteins%40net.bio.net (by jin.zongwen from gmail.com)
Fri Jan 15 01:33:25 EST 2010


I recently started to purify a protein containing zinc finger modif.

the protein consist of flexible linker and his tag in the N term and C
terminal of the mofit.

the Zinc finger itself is consisted from three zinc finger motifs.

It expressed well in the E.coli, and I was able to purify the protein, but
when I tried to concentrate it by using a membrane filter( amicon centricon)
the protein start to aggregate.

what is more a concern is that gentle swiring in 4'C would generate cotton
like aggregates in the tube.

I would like to ask if anyone have expperience on zinc finger protein,
expecially small zinc finger proteins like this one.(15kDa)

any suggestion are welcomed.

Best regards to every one

-- 
Zongwen Jin
Biomolecular Engineering Lab
Dept. of Biological Sciences, KAIST
335 Gwahangno, Guseng-dong, 305-701
Yuseong-gu, Daejeon, Republic of Korea
Researcher ID: B-4590-2009
Lab: +82-42-350-2656
Mobile: +82-10-7460-9518
E-mail: sutem from kaist.ac.kr
K-Internet : 070-8288--0606


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