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Identification of phage recombinase function unit in genus Corynebacterium
Yizhao Chang1, Qian Wang2, Tianyuan Su3
1State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong, China.
Applied Microbiology and Biotechnology
|June 16, 2021
Summary
We identified 60 novel phage recombinase function units (PRFUs) in Corynebacterium genomes. A PRFU from C. aurimucosum showed high efficiency for recombineering, advancing genetic engineering in this genus.
Area of Science:
- Microbiology
- Molecular Biology
- Genomics
Background:
- Phage recombinase function units (PRFUs) are essential for phage life cycles and have been repurposed for recombineering in Escherichia coli.
- Most PRFUs function optimally only in their native hosts, necessitating the discovery of new PRFUs for diverse species.
Purpose of the Study:
- To identify and characterize novel PRFUs within the Corynebacterium genus.
- To evaluate the recombineering efficiency of identified PRFUs in Corynebacterium glutamicum.
Main Methods:
- Bioinformatic analysis of 423 Corynebacterium genomes using a database-to-database search method to predict PRFUs.
- Experimental verification of recombineering capabilities for eight representative PRFUs in Corynebacterium glutamicum ATCC 13032.
Main Results:
- Identified 60 unique PRFUs across 8 distinct evolutionary types within the Corynebacterium genus.
- Experimentally confirmed the recombineering ability of 8 representative PRFUs in C. glutamicum.
- The PRFU from Corynebacterium aurimucosum demonstrated the highest efficiency (10^-2 magnitude) for both single-stranded and double-stranded DNA-mediated recombineering.
Conclusions:
- This study presents the first systematic bioinformatic mining and analysis of PRFUs in Corynebacterium.
- The identified PRFUs, particularly the highly efficient one from C. aurimucosum, are valuable tools for advancing genome engineering in the Corynebacterium genus.
- These findings offer new insights into the study and application of PRFUs in microbial genetics.
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