Vectors for enhanced gene expression and protein purification in Salmonella.
Improved expression vectors have been tested for protein expression studies in Salmonella spp. They are derived from the broad host range expression vector pNSGroE [Seleem, M.N., Vemulapalli, R., Boyle, S.M., Schurig, G.G. and Sriranganathan, N., 2004. Improved expression vector for Brucella species. Biotechniques 37, 740-744] and have several advantages (i) small in size (less than 3 kb); (ii) possess eleven unique restriction site to facilitate directional cloning; (iii) express proteins as His-tagged fusions for easy detection and purification; (iv) carry different promoters for various level of expression and (v) carry an UP element and RNA stem-loop for enhanced gene expression. We have demonstrated the ability of the new vectors to stably express heterologous proteins in Salmonella. We also demonstrated the utility of our vectors by detecting expression and purification of recombinant GFP. Our results suggest that these new vectors should improve gene expression in Salmonella, particularly those aimed at foreign antigen delivery.
Seleem
Mohamed N
MN
Institute for Critical Technology and Applied Science, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Ali
Mohammed
M
Boyle
Stephen M
SM
Sriranganathan
Nammalwar
N
D005786:Gene Expression Regulation; D005822:Genetic Vectors; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification*; D050436:Regulatory Elements, Transcriptional; D012475:Salmonella / Q000235:genetics*; D014158:Transcription, Genetic
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins