Large inserts for big data: artificial chromosomes in the genomic era

Area of Science:

• Microbial genomics and bioinformatics
• Natural product discovery
• Synthetic biology and metabolic engineering

Background:

• Vast numbers of microbial genomes are available, revealing extensive genetic potential for specialized bioactive compounds.
• Most biosynthetic gene clusters (BGCs) encoding these compounds are cryptic, remaining unexpressed under standard laboratory conditions.

Purpose of the Study:

• To provide an overview of vectors and cloning systems designed for large biosynthetic gene clusters (BGCs).
• To highlight successful strategies for heterologous expression of BGCs in suitable host organisms.

Main Methods:

• Review of established and novel cloning vectors capable of harboring large DNA fragments.
• Analysis of different approaches for transferring and expressing entire BGCs in heterologous hosts.
• Compilation of successful case studies demonstrating heterologous expression of microbial BGCs.

Main Results:

• Development of diverse cloning systems and vectors enabling the manipulation of large DNA fragments.
• Successful transfer and expression of previously cryptic BGCs in various heterologous hosts.
• Demonstration of the feasibility of accessing novel bioactive compounds through heterologous expression.

Conclusions:

• Heterologous expression is a powerful strategy to unlock the biosynthetic potential of cryptic BGCs in microbial genomes.
• Advances in cloning technologies and vectors are crucial for the successful transfer and expression of large BGCs.
• This approach significantly expands the repertoire of natural products accessible for drug discovery and biotechnology.