Activation of Silent Gene Clusters Mediated by Promoter Substitution

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Promoter substitution is using a new promoter to replace the original promoter of a silenced gene, thus allowing normal expression of the silenced gene in the original host. Since the secondary metabolites of fungi may be toxic to the original host, researchers often use inducible promoters to replace the original promoters of silenced genes to not affect the fungus's growth and activate the silenced genes to obtain new active natural products. However, when performing promoter substitution, the promoters of all silenced biosynthetic genes in the gene cluster need to be replaced, and the workload is high. Lifeasible is committed to developing promoter substitution-mediated activation strategies for silent gene clusters to help customers achieve efficient utilization of fungal resources.

Figure 1. Physical map of the epothilone biosynthetic gene cluster and the promoter Pepo. (Van Dijck P, et al., 2017)

Biosynthetic gene promoter substitution. We use inducible promoters to replace the original promoters of silenced biosynthetic genes and then drive the expression of the target genes in the presence of inducers to synthesize new metabolites.
Transcriptional regulatory gene promoter substitution. Fungal biosynthetic gene clusters usually contain multiple transcriptional regulatory genes, which are primarily responsible for synthesizing transcriptional regulators that then specifically bind to the promoter sequences of other genes in the gene cluster, thereby activating the expression of a specific gene cluster.

The specific services we can provide are as follows. Including but not limited to.

Promoter cloning	Known promoter sequences Unknown promoter sequences
Promoter activity analysis	Report genetic test
Analysis of promoter core elements	Promoter 2.0 Prediction Server Softberry Plant CARE
Genetic editing	CRISPR/Cas9
Dual fluorescence reporting system	Analysis of promoter structure Promoter SNP analysis Interaction analysis of transcription factors and promoters

The commonly used eukaryotic promoters are as follows.

Eukaryotic promoters	Types of transcribed RNA	Expression features	Other features
CMV	mRNA	Constitutive expression	May contain an enhancer that will silence in some cell types
EF1a	mRNA	Constitutive expression	Stable expression in all cell types and physiological states
SV40	mRNA	Constitutive expression	May contain an enhancer
CAG	mRNA	Constitutive expression	Contains CMV enhancer
UAS	mRNA	Specific expression	Requires the presence of the Gal4 gene to activate the promoter
Ac5	mRNA	Constitutive expression	Expression systems commonly used in Drosophila
TEF1	mRNA	Constitutive expression	Similar to the mammalian EF1α promoter
GDS	mRNA	Constitutive expression	Great ability to start expressions
ADH1	mRNA	Inhibition with ethanol	This promoter has a strong initiation ability and high expression level when it is full-length; it is constitutively expressed when it is truncated and has a low expression level
…	…	…	…

Lifeasible can provide full operational services for promoter substitution-mediated activation of silent gene clusters, helping our customers to reduce project time. As your trusted partner, we can meet all your fungal phylogenetic analysis needs and provide efficient, high-quality services. If you want to know the details, please contact us.

Reference

Yue XJ, et al. Effects of transcriptional mode on promoter substitution and tandem engineering for the production of epothilones in Myxococcus xanthus. Appl Microbiol Biotechnol. 2018 Jul; 102(13): 5599-5610.

For research or industrial raw materials, not for personal medical use!

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