PHT253

The pHT253 vector contains the coding sequence for an 8xHis tag and allows high-level intracellular production of recombinant N-terminal His 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). pHT253 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
• N-terminal 8xHis-tag
• E. coli / B. subtilis shuttle vector

The pHT253 vector is available from MoBiTec GmbH (Göttingen)

References

Hoang Duc Nguyen, Trang Thi Phuong Phan, Wolfgang Schumann
Expression vectors for the rapid purification of recombinant proteins in Bacillus subtilis.
Curr Microbiol: 2007, 55(2);89-93
[PubMed:17624574] [WorldCat.org] [DOI] (P 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)

Hoang Duc Nguyen, Quynh Anh Nguyen, Rita C Ferreira, Luis C S Ferreira, Linh Thuoc Tran, Wolfgang Schumann
Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability.
Plasmid: 2005, 54(3);241-8
[PubMed:16005967] [WorldCat.org] [DOI] (P p)