Gene Conditional Expression for Bacteria

Controlled (or conditional) gene expression provides a mean to generate conditional defects, which is essential for gene function validation. Creative Biolabs provides a lot of controlled gene expression tools to identify and validate the essential genes for the pathogenic bacteria.

Controlled gene expression involves the replacement or disruption of the wild-type promoter by the insertion of an inducible promoter immediately upstream of the gene of interest. Gene expression is activated by a small molecule (inducer) that turns on promoter activity. If the test gene is vital, the strain depends on the inducer for living and growth. The activities of some inducible promoters can also be tested by changing inducer concentrations allowing the analysis of the response of the strain under semi-permissive conditions in which the concentration of the target protein is just sufficient to enable growth.

Figure 1. TET system controlled gene expression

Creative Biolabs can provide different gene conditional expression tools for both Gram-negative and Gram-positive species. These inducible promoters include that regulated by arabinose, rhamnose, tetracycline, lactose (IPTG), xylose, and fucose. As long as the genomic sequence is available, gene essentiality can be identified and assessed. Our platform demonstrates both the reproducibility and reliability for determining gene essentiality and is extremely suitable for systematic and large-scale essential gene identification and validation in diploid pathogenic microorganisms.

After identifying essential genes by conditional expression method, searching for orthologous genes in the human and other bacterial genomes can further aid drug discovery. Those genes absent in humans but conserved in pathogens may represent broad spectrum targets. The specific genes with no human or other bacteria counterpart may be suitable as narrow spectrum targets.

In addition to identifying target genes, constructing a comprehensive gene conditional expression mutant strain collection has many applications which can significantly accelerate functional annotation. The other useful applications for conditional mutant strains may include:

• Serve as sensitized whole cell assays suitable for high throughput drug screening a wide spectrum of preferred targets regardless of whether the functional or biochemical information is available.
• Involved in a wide range of cellular processes, including growth assay, biofilm formation and in vivo screens to identify virulent-related factors.
• Apply to transcriptional profiling for essential drug targets.

Creative Biolabs is a trusted partner in the aspect of gene conditional expression. Our one-stop service is time-saving, high-quality, tailor-made, and economical. For more detailed information, please feel free to contact us or directly sent us an inquiry.

Reference

1. Lynn Miesel, Jonathan Greene & Todd A. Black. (2003). “Microbial genetics: Genetic strategies for antibacterial drug discovery”. Nature Reviews Genetics 4, 442-456 (June 2003).

For Research Use Only.