解淀粉芽孢杆菌XH7的全基因组转录图谱及基于RNA测序数据的强启动子高通量筛选

The global transcriptional landscape of Bacillus amyloliquefaciens XH7 and high-throughput screening of strong promoters based on RNA-seq data.

作者信息

Liao Yuling, Huang Lianggang, Wang Bin, Zhou Feng, Pan Li

机构信息

School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong 510006, China; Star Lake Bioscience Co., Inc., Zhaoqing, Guangdong 526060, China.

School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, Guangdong 510006, China.

出版信息

Gene. 2015 Oct 25;571(2):252-62. doi: 10.1016/j.gene.2015.06.066. Epub 2015 Jun 28.

Abstract

Bacillus amyloliquefaciens is an important industrial microbe for the production of many industrial enzymes and primary metabolites. Although the complete genome sequence of B. amyloliquefaciens has been now published, transcript structures of B. amyloliquefaciens remain poorly defined. In this study, high-throughput RNA sequencing (RNA-seq) technology was applied to dissect the transcriptome of B. amyloliquefaciens strain XH7. In total, 3936 out of a total of 4204 B. amyloliquefaciens genes (93.6%) were transcribed under the selected growth condition. Transcriptional start sites (TSS) of 1064 annotated genes and 749 operons were identified. To screen for strong promoters, a beta-galactoside reporter was fused to eight candidate promoters from 288 genes with higher expression levels (RPKM values) than the control gene P43-bgaB. The results illustrated that the candidate promoter Pr2 (promoter for the sigW gene) displayed the strongest beta-galactosidase specific activity during the post-log phase, suggesting that it could be used effectively for heterologous gene expression. The presented data will contribute to the further study of the B. amyloliquefaciens transcriptome by identifying useful promoters for industrial uses.

摘要

解淀粉芽孢杆菌是一种重要的工业微生物,可用于生产多种工业酶和初级代谢产物。尽管解淀粉芽孢杆菌的完整基因组序列现已公布,但其转录结构仍不清楚。在本研究中,采用高通量RNA测序(RNA-seq)技术剖析了解淀粉芽孢杆菌XH7菌株的转录组。在选定的生长条件下,总共4204个解淀粉芽孢杆菌基因中有3936个(93.6%)被转录。鉴定了1064个注释基因和749个操纵子的转录起始位点(TSS)。为了筛选强启动子,将β-半乳糖苷报告基因与来自288个表达水平(RPKM值)高于对照基因P43-bgaB的基因的八个候选启动子融合。结果表明,候选启动子Pr2(sigW基因的启动子)在对数后期显示出最强的β-半乳糖苷酶比活性,表明它可有效地用于异源基因表达。所提供的数据将通过鉴定工业用途的有用启动子,有助于进一步研究解淀粉芽孢杆菌的转录组。

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