A general method for the consecutive integration of single copies of a heterologous gene at multiple locations in the Bacillus subtilis chromosome by replacement recombination

Article Abstract:

A method based on the transformability of Bacillus subtilis strains allows the integration of single copies of a heterologous gene at multiple locations in the bacterial chromosome. A thymine-auxotrophic B. subtilis mutant is transformed to prototrophy by integrating a gene at the required position. The mutant is then transformed with a DNA fragment carrying the heterologous gene. A repetition of the procedures allows the isolation of strains containing multiple copies of the heterologous gene at different positions. The method does not require the simultaneous integration of markers.

Methods, Microbial genetic engineering

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Functional analysis of the secretory precursor processing machinery of Bacillus subtilis: identification of a eubacterial homolog of archaeal and eukaryotic signal peptidases

Article Abstract:

The sip gene family of Bacillus subtilis has been studied and a eubacterial homolog of archaeal and eukaryotic signal peptidases identified. Of the five chromosomal genes, SipS and SipT are of great importance for protein secretion. The newly identified SPase SipW is very similar to SPases from archaea and the endoplasmic reticular (ER) membrane of eukaryotes. That would indicate that the enzymes make up a subfamily, type I SPases, conserved in all three domains.

Research, Analysis, Endoplasmic reticulum, Proteases, Proteins, Protein synthesis, Signal peptides

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Phage display of an intracellular carboxylesterase of Bacillus subtilis: Comparison of Sec and Tat pathway export capabilities

Article Abstract:

The development of the phage display technology method is described for fusion of an intracellular carboxylesterase of Bacillus subtilis to the phage minor coat protein g3p. The effects of Sec dependent and Tat dependent signal peptides were assessed, on the translocation of carboxylesterase for optimizing the phage display of this enzyme normally restricted to the cytoplasm.

Esterases, Chemical properties, Carboxyl compounds

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