Abstract:
:The major immunodominant integral outer membrane protein C (OmpC) from Salmonella typhi Ty21a was overexpressed, without the signal peptide, in Escherichia coli. The protein aggregates as inclusion bodies (IBs) in the cytoplasm. OmpC from IBs was solubilized with 4 M urea and refolded. This involved rapid dilution of unfolded OmpC into a refolding buffer containing polyoxyethylene-9-lauryl ether (C(12)E(9)) and glycerol. The refolded OmpC (rfOmpC) was shown to be structurally similar to the native OmpC by SDS-PAGE, Western blotting, tryptic digestion, ultrafiltration, circular dichroism, and fluorescence spectroscopic techniques. Crystals of rfOmpC were obtained in preliminary crystallization trials. The rfOmpC also sets a stage for rational design by recombinant DNA technology for vaccine design and high resolution structure determination.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Kumar PD,Krishnaswamy Sdoi
10.1016/j.pep.2004.12.023keywords:
subject
Has Abstractpub_date
2005-03-01 00:00:00pages
126-33issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(04)00444-9journal_volume
40pub_type
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