期刊
JOURNAL OF BIOTECHNOLOGY
卷 149, 期 1-2, 页码 16-20出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2010.06.021
关键词
Bacillus thuringiensis cry3Aa; Expression system; Promoter modification; Bacillus subtilis
资金
- Ministry of Education, Science and Technology
- Ministry of Knowledge Economy of Korea [DTC0050812]
Bacillus thuringiensis produces crystal proteins (Cry) that account for up to 25% of the dry cell weight during the stationary phase. The high-level expression and stationary phase-specific autoinduction of the cry gene led to development of a cry promoter-based Bacillus expression system. Among the various cry promoters, cry3Aa promoter was selected by comparing the lacZ expression levels in Bacillus subtilis. An extracellular enzyme cellulase was highly upregulated during the stationary phase while under control of the cry3Aa promoter. Improvement of the cry3Aa promoter was obtained by modification of the promoter sequence. Specifically, a 5-fold increase in lacZ expression was obtained by changing both the -35 and - 10 boxes of the cry3Aa promoter to the consensus sequence of the sigma(A)-dependent promoter of B. subtilis. The modified cry3Aa promoter produced a significantly higher yield of AprE, which suggests that the promoter may be useful for high-level protein expression in B. subtilis. (C) 2010 Elsevier B.V. All rights reserved.
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