Bacillus subtilis pBacTag Tagging Vector, 5 µg

The pBacTag Vectors are Bacillus subtilis epitope- and fluorescence-tagging integration vectors for the directed functional analysis of B. subtilis genes.

All pBacTag tagging vectors are derivatives of pMutin vectors. pBacTag tagging vectors can replicate in E. coli but are unable to propagate in B. subtilis, enabling chromosomal integration with B. subtilis (and other bacterial species in which pBR322 based plasmids are not able to replicate). The IPTG inducible Pspac promoter allows controlled expression of genes that are located downstream of the target gene.

For proper cloning, the vectors contain a multiple cloning site downstream of the Pspac with the unique restriction sites KpnI, Eco47III, ClaI and EagI. To ensure efficient termination of transcription of the hybrid gene, the vectors contain the trpA terminator of the E. coli tryptophan operon downstream of the tag.

Tagging or inactivation of target genes is achieved by chromosomal integration of a pBacTag Vector via homologous recombination. The gene of interest can be selectively inactivated within the chromosome, enabling phenotypic studies of the resulting mutant. Alternatively, the gene of interest can be expressed chromosomally as a translational fusion protein with an N-terminal epitope or localization tag. After inactivation or tagging, genes localized directly downstream of the gene of interest can be controlled by an IPTG inducible promoter to ensure their expression. 

Six pBacTag tagging vectors are offered as either epitope tags or localization tags: 

• Epitope tags (PBT001, PBT002, PBT003) - used for selective purification of fusion proteins and for their detection by commercially available antibodies. Since the tags are very short, protein function is usually not disturbed.

• Localization tags (PBT004, PBT005, PBT006) - helpful for localization studies of target proteins. The fusion proteins that contain a fluorescing tag can be analyzed for their cell compartmental localization. The GFP+ tag, pBacTagGFP+ (PBT004), is an improved variant of the common GFP, which produces enhanced fluorescence.