Expression, Purification, Refolding, and Characterization of a Neverland Protein From Caenorhabditis elegans.

ABSTRACT: Steroid hormones that serve as vital compounds are necessary for the development and metabolism of a variety of organisms. The neverland (NVD) family genes encode the conserved Rieske-type oxygenases, which are accountable for the dehydrogenation during the synthesis and regulation of steroid hormones. However, the His-tagged NVD protein from Caenorhabditis elegans expresses as inclusion bodies in Escherichia coli BL21 (DE3). This bottleneck can be solved through refolding by urea or the introduction of a maltose-binding protein (MBP) tag at the N-terminus. Through further research on purification after the introduction of a MBP tag at the N-terminus, the CD measurement and fluorescence-based thermal shift assay indicated that MBP was favorable for the NV

SUBMITTER: Mao S 

PROVIDER: S-EPMC7609953 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature