Heterologous expression of gene clusters

Gene clusters that have been cloned from their native host can be expressed heterologously in a different host organism. This can help improve production of known products, or aid the discovery of novel natural products.

Cloned gene clusters can be introduced into a host strain via intergenic conjugation.

Useful host strains:

• Streptomyces albus [1]

This species has a naturally minimised genome and fast growth, which can help improve production of a target gene cluster.

Streptomyces superhosts [2]:

• Streptomyces coelicolor M1146

Δact red cpk cda

• Streptomyces coelicolor M1152

Δact red cpk cda rpoB[C1298T] rpsL [A262G C271T] These Streptomyces superhosts were constructed from Streptomyces coelicolor M145. Four exogenous secondary metoblite gene clusters were deleted (actinorhodin, prodiginine, CPK and CDA) to remove competition of carbon and nitrogen resources. For M1152, point mutations were introduced into rpoB and rpsL to help increase production of biosynthetic gene clusters.

[1] Zaburannyi, N. et al. (2014) ‘Insights into naturally minimised Streptomyces albus J1074 genome’ BMC genomics 15:97

[2] Gomez-Escribano, J. P. and Bibb, M. J. (2011) ‘Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters’ Microbial Biotechnology, 4(2):207-15. doi: 10.1111/j.1751-7915.2010.00219.x.