PHT253

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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

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)