Yeast-Based Assembly of Phage Genomes

Propagation of phage genomes in bacterial hosts can be toxic to the host, thus limiting the efficiency of phage genome engineering using methods such as homologous recombination, BRED, and in vivo recombineering. This problem can be overcome by using Saccharomyces cerevisiae rather than bacteria as an intermediate host for genetic manipulation. Homologous recombination is particularly efficient in Saccharomyces cerevisiae, and the phage genome does not cause toxicity in yeast and can be stably maintained. Using this method, phage genomes were captured in a Saccharomyces cerevisiae-bacteria shuttle vector. Phage genomes assembled, modified, and propagated in yeast have been isolated and introduced into bacteria to generate functional phage particles. This technology has been used to capture and genetically modify the genomes of E. coli phages T3 (38,208 bp) and T7 (39,937 bp) and Klebsiella phage K11 (41,181 bp). It was further used to capture and archive the genome of the fully reconstituted phage φX174 (6,302 bp).

Fig 1 Yeast-based phage assembly.(Gibb B, et al. 2021)

Our Services

Creative Biolabs has been committed to phage engineering for a long time and has accumulated rich experience in the development of different engineered phages such as reporting phages, labeled phages, therapeutic phages, etc. Our services are listed as follows, but are not limited to:

• Designing a decompressed phage
• Building phage genomes in yeast
• Transplantation of wild-type phage to and from yeast
• Assembly and recovery of a decompressed phage
• Initial phenotypic characterization of phage
• Lysis curves analysis of phage
• DNA design and manipulation and sequencing

Our Advantages

• Years of experience in the yeast-based assembly of bacteriophage genomes.
• Combining different omics approaches such as transcriptomics and proteomics with recombinant DNA technology.
• Advanced yeast culture technology.
• Optimize media selection, biomass control and product induction.
• Cost-effective and high-quality products.

Project Workflow

• Customer design advisory
• Phage project discussion
• Building phage genomes in yeast service
• Phenotypic characterization of phage
• Phage plaque assays
• Assessment of phage lysis efficiency
• Evaluation of the novel phage
• Project delivery

Highlight of Yeast-Based Assembly of Bacteriophage Genomes

• Methods based on yeast or in vitro phage genome assembly have been developed to avoid the potentially toxic effects of phage replication on bacterial hosts.
• Since yeast cells have an efficient recombination system, and phage genes are nontoxic to yeast.

Creative Biolabs can meet the needs of customers by providing yeast-based assembly of bacteriophage genomes services on time and on budget. We have in-depth knowledge and experience of the tools and processes involved in the phage projects. Our skilled and dedicated scientific researchers ensure that the most suitable methods and techniques are selected for your project. If necessary, please feel free to contact us.

Reference

• Gibb B, Hyman P, Schneider C L. The Many Applications of Engineered Bacteriophages - An Overview[J]. Pharmaceuticals, 2021, 14(7): 634.

Please kindly note that our services can only be used to support research purposes (Not for clinical use).