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Promiscuous Trans-splicing Activities Revealed by Next Generation Sequencing-based Analysis of 298 Split Inteins

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  • Protein Engineering
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Abstract

Protein trans-splicing is a naturally occurring process in which two protein fragments are ligated by a reaction between two intein domains, called split inteins. Despite their usefulness in research, the reactivity and structure of only a few split inteins have been studied. We used cell-based kanamycin selection and next-generation sequencing (NGS) to simultaneously measure the splicing reactivity of 298 N-intein–C-intein combinations derived from the DnaE gene of cyanobacteria. Additionally, we confirmed the splicing activities by measuring the growth of cells individually harboring each split intein under kanamycin selection. Overall, the N-intein–C-intein combinations were promiscuous in their trans-splicing activities, although certain combinations did not splice actively. These results and the NGS-based analysis in this research would be helpful for the development of novel split inteins and further understanding of the trans-splicing mechanism.

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Acknowledgments

We thank the members of the Bang Lab for their critical comments during this work. This work was supported by: (i) the Mid-career Researcher Program (NRF-2018R1A2A 1A05079172) from the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT & Planning; (ii) the Bio & Medical Technology Development Program of the NRF, funded by the Korean government (MSIT; NRF-2016M3A9B6948494); (iii) the Bio & Medical Technology Development Program of the NRF, funded by the Korean government (MSIT; NRF-2018M3A9H3024850) and (iV) the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI18C2282).

The authors declare no conflict of interest.

Neither ethical approval nor informed consent was required for this study.

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  1. Department of Chemistry, Yonsei University, Seoul, 03722, Korea

    Han Na Seo & Duhee Bang

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  1. Han Na Seo
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  2. Duhee Bang
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Correspondence to Duhee Bang.

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Aptamer-based Fluorescent Assay for Simple and Sensitive Detection of Fipronil in Liquid EggsPromiscuous Trans-splicing Activities Revealed by Next Generation Sequencing-based Analysis of 298 Split Inteins

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Seo, H.N., Bang, D. Promiscuous Trans-splicing Activities Revealed by Next Generation Sequencing-based Analysis of 298 Split Inteins. Biotechnol Bioproc E 25, 293–301 (2020). https://doi.org/10.1007/s12257-019-0394-x

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  • DOI: https://doi.org/10.1007/s12257-019-0394-x

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