A dual protein expression system in Bacillus subtilis

Annie Y. Chan, Mei M. Chan, Hei M. Lo, Yun Chung Leung, Boon L. Lim

Research output: Journal article publicationJournal articleAcademic researchpeer-review

14 Citations (Scopus)

Abstract

We have developed a dual expression system for the simultaneous overexpression of two proteins in Bacillus subtilis. Two candidate genes, xylanase (xynA) and glucanase (bglS) from B. subtilis strain 168, which were engineered with independent Shine-Dalgarno (SD) sequences, were cloned in tandem into a transfer vector, which was then transformed into B. subtilis strain 1A304 (φ105MU331). The genes were under the transcriptional control of a strong promoter of a bacteriophage, φ105, where transcription was initiated upon thermal induction. Six constructs were made to compare the factors that affected the yields of the gene products. The expression level of each candidate gene was found to correspond to its position relative to the phage promoter, irrespective of the identity of the insert. The lower expression level of the second insert might have been due to limited resources for protein synthesis, a short half-life of the mRNA, or an early termination of the RNA polymerase. Curiously, gene duplications in tandem did not lead to further increase in production.
Original languageEnglish
Pages (from-to)337-342
Number of pages6
JournalProtein Expression and Purification
Volume26
Issue number3
DOIs
Publication statusPublished - 1 Dec 2002

Keywords

  • Bacillus subtilis
  • Dual expression
  • Glucanase
  • Prophage
  • Xylanase

ASJC Scopus subject areas

  • Biotechnology

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