Over-expression of proteins using a modified pBAD24 vector in E. coli expression system

Biotechnol Lett. 2009 Jul;31(7):1031-6. doi: 10.1007/s10529-009-9976-6. Epub 2009 Mar 29.

Abstract

A modified pBAD24 vector (pBAD24M) was constructed with the araBAD promoter of the arabinose operon along with T7g10 sequence elements and a modified Shine-Dalgarno sequence. While both green fluorescent protein and granulocyte colony stimulating factor showed negligible expression under the original pBAD24 vector, they were expressed at >35% of total cellular protein with the modified vector. Similar results were obtained for staphylokinase wherein the pBAD24-SAK construct yielded 8 ng/10(6) c.f.u. of E. coli induced cells while the pBAD24M-SAK vector showed nearly 55 ng/10(6) c.f.u. induced bacterial cells as tested by ELISA. Interestingly, the expression levels using modified pBAD24 vector matched that achieved with T7 promoter based vector system. The modified pBAD24 vector therefore represents a simple and a useful prokaryotic expression system for efficient repression, modulation and elevated protein expression levels.

MeSH terms

  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Gene Expression*
  • Genes, Bacterial
  • Genes, Viral
  • Genetic Vectors*
  • Podoviridae / genetics
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics

Substances

  • Recombinant Proteins