Insoluble Protein

Zhongguo Xiong xiong at mozart.biosci.arizona.edu
Mon Oct 18 15:51:00 EST 1993


Dear Netters:

I have recently encountered a insoluble recombinant protein and need all kinds
of suggestions or comments I can get. I over-expressed a plant RNA viral
protein (possible a RNA-depedent RNA polymerase protein) in E. coli. The
protein formed an inclusion body in the bacteria. I had no problems in
solublizing the protein with urea and the subsequent protein purification in
the nickel column. Then I dialyzed the purified protein against a series of
Urea buffers with decreasing concentrations of Urea (from 8M to 0). Inspite of
the precaution, the protein still formed precipitates. I tried to resolublize
the protein in 4 M urea and dit not see any protein in SDS PAGE gel.

Are there any way to make the protein soluble in a non-denaturing condition?
Any comments and suggestions are welcome.


Z. Xiong
Department of Plant Pathology
University of Arizona, Tucson, AZ 85721
Xiong at biosci.arizona.edu  



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