Control of total GFP expression by alterations to the 3' region nucleotide sequence.

ABSTRACT:

Background

Previously, we distinguished the Escherichia coli type II cytoplasmic membrane translocation pathways of Tat, Yid, and Sec for unfolded and folded soluble target proteins. The translocation of folded protein to the periplasm for soluble expression via the Tat pathway was controlled by an N-terminal hydrophilic leader sequence. In this study, we investigated the effect of the hydrophilic C-terminal end and its nucleotide sequence on total and soluble protein expression.

Results

The native hydrophilic C-terminal end of GFP was obtained by deleting the C-terminal peptide LeuGlu-6×His, derived from pET22b(+). The corresponding clones induced total and soluble GFP expression that was either slightly increased or dramatically reduced, apparently through reconstruction o

SUBMITTER: Lee SJ 

PROVIDER: S-EPMC3726318 | biostudies-literature | 2013 Jul

REPOSITORIES: biostudies-literature