PHT255

From SubtiWiki
Revision as of 10:25, 28 August 2017 by Bzhu (talk | contribs)
Jump to: navigation, search

Disclaimer

The content of this page/section is provided by a third party. SubtiWiki acknowledges that this third party content is obtained from a source believed to be reliable. Please note that no guarantees are made by SubtiWiki as to reliability of this third party information. Provision of this third party content is for general information purpose and does not constitute a recommendation. SubtiWiki does not own copyright over this third party content (including but not limited to text, images, and media).

PHT255.PNG PHT255-Legende.jpg

The pHT255 vector contains the coding sequence for a Strep tag and allows high-level intracellular production of recombinant C-terminal Strep tagged protein with B. subtilis. The tag can be used for detection or purification of the produced protein of interest. The expression is controlled by the strong Pgrac100 promoter, consisting of the groeESL promoter of B. subtilis with improved regulatory elements fused to the lac operator allowing their induction by IPTG. Nucleotides were optimized at the conserved regions of the groESL promoter including the UP element, the −35 and the −15 region. Combination of these changes into one promoter enhanced the amount of recombinant proteins accumulating intracellularly up to about 30% of the total cellular protein of B. subtilis (Phan et al., 2011). pHT255 is an E. coli / B. subtilis shuttle vector, that provides ampicillin resistance to E.coli and chloramphenicol resistance to B. subtilis.

Features

  • Strong promoter with improved regulatory elements
  • Enhanced amount of produced recombinant proteins
  • IPTG inducible gene expression
  • C-terminal Strep tag
  • E. coli / B. subtilis shuttle vector

The pHT255 vector is available from MoBiTec GmbH (Göttingen)

References

Trang Thi Phuong Phan, Linh Thuoc Tran, Wolfgang Schumann, Hoang Duc Nguyen
Development of Pgrac100-based expression vectors allowing high protein production levels in Bacillus subtilis and relatively low basal expression in Escherichia coli.
Microb. Cell Fact.: 2015, 14;72
[PubMed:25990516] [WorldCat.org] [DOI] (I e)

Trang Thi Phuong Phan, Hoang Duc Nguyen, Wolfgang Schumann
Development of a strong intracellular expression system for Bacillus subtilis by optimizing promoter elements.
J. Biotechnol.: 2012, 157(1);167-72
[PubMed:22100269] [WorldCat.org] [DOI] (I p)

Trang Thi Phuong Phan, Hoang Duc Nguyen, Wolfgang Schumann
Novel plasmid-based expression vectors for intra- and extracellular production of recombinant proteins in Bacillus subtilis.
Protein Expr. Purif.: 2006, 46(2);189-95
[PubMed:16125412] [WorldCat.org] [DOI] (P p)