Скачать или смотреть Paul D'Agostino | Applying DiPaC towards the capture and expression of biosynthetic gene clusters

Paul D'Agostino - Dresden University of Technology, Germany

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Applying Direct Pathway Cloning (DiPaC) towards the capture and expression of cyanobacterial biosynthetic gene clusters

Microbial natural products are a vital source of inspiration for drug development. Therefore, the discovery of novel and bioactive natural products is essential to combat emerging new pathogens and to counteract the exacerbating crisis of microbial resistance. While cyanobacteria have proven to be an excellent source of structurally intriguing and bioactive natural products, several factors impede the efficient discovery and exploitation of these molecules including slow growth rates, challenging or ineffective genetic manipulation techniques and low molecule titres. To overcome these difficulties and speed-up discovery rates, we recently developed a synthetic biology cloning strategy termed Direct Pathway Cloning (DiPaC) for the capture and heterologous expression of small- to mid-sized cyanobacterial gene clusters. DiPaC utilises long-range PCR and in vitro DNA assembly to facilitate pathway assembly, cluster refactoring and direct transfer into any vector backbone in vitro.

In the first part of my talk, I will describe our application of DiPaC towards the experimental validation of the hapalosin and cyanobacterin biosynthetic gene clusters. In the second part of my talk, I will provide an outline for our plans to target cryptic biosynthetic gene clusters for the discovery of novel molecules from a range of symbiotic cyanobacteria, particularly those isolated from lichens or cycads.

Due to its simplicity, we envisage DiPaC to become an essential method for biosynthetic gene cluster capture and metabolic pathway construction with significant applications for the functional characterisation of novel natural product pathways well into the future.

This talk is part of the CyanoWorld online seminar series, hosted by Nicolas Schmelling and Ilka Axmann, and was held on Zoom on August 26th, 2021.

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