基于地衣芽孢杆菌转录组鉴定强启动子

Identification of strong promoters based on the transcriptome of Bacillus licheniformis.

作者信息

Liu Xin, Yang Haiyan, Zheng Junwei, Ye Yanrui, Pan Li

机构信息

School of Bioscience and Bioengineering, Guangzhou Higher Education Mega Center, South China University of Technology, Guangzhou, 510006, Guangdong Province, People's Republic of China.

出版信息

Biotechnol Lett. 2017 Jun;39(6):873-881. doi: 10.1007/s10529-017-2304-7. Epub 2017 Feb 25.

DOI:10.1007/s10529-017-2304-7

PMID:28238059

Abstract

OBJECTIVES

To expand the repertoire of strong promoters for high level expression of proteins based on the transcriptome of Bacillus licheniformis.

RESULTS

The transcriptome of B. licheniformis ATCC14580 grown to the early stationary phase was analyzed and the top 10 highly expressed genes/operons out of the 3959 genes and 1249 operons identified were chosen for study promoter activity. Using beta-galactosidase gene as a reporter, the candidate promoter pBL9 exhibited the strongest activity which was comparable to that of the widely used strong promoter p43. Furthermore, the pro-transglutaminase from Streptomyces mobaraensis (pro-MTG) was expressed under the control of promoter pBL9 and the activity of pro-MTG reached 82 U/ml after 36 h, which is 23% higher than that of promoter p43 (66.8 U/ml).

CONCLUSION

In our analyses of the transcriptome of B. licheniformis, we have identified a strong promoter pBL9, which could be adapted for high level expression of proteins in the host Bacillus subtilis.

摘要

目的

基于地衣芽孢杆菌的转录组，拓展用于蛋白质高水平表达的强启动子库。

结果

分析了生长至早期稳定期的地衣芽孢杆菌ATCC14580的转录组，从鉴定出的3959个基因和1249个操纵子中挑选出前10个高表达基因/操纵子用于研究启动子活性。以β-半乳糖苷酶基因作为报告基因，候选启动子pBL9表现出最强的活性，与广泛使用的强启动子p43相当。此外，来自茂原链霉菌的谷氨酰胺转胺酶原（pro-MTG）在启动子pBL9的控制下表达，36小时后pro-MTG的活性达到82 U/ml，比启动子p43（66.8 U/ml）高23%。