An improved molecular tool for screening bacterial colonies using GFP expression enhanced by a Dictyostelium sequence

Kondo, T.; Yumura, S.

doi:10.1101/821157

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bioRxiv · 10.1101/821157

An improved molecular tool for screening bacterial colonies using GFP expression enhanced by a Dictyostelium sequence

Kondo, T.·Yumura, S.

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Abstract

During molecular cloning, screening of bacterial transformants is a time-consuming and labor-intensive process. However, tractable tools that can be applied to various vectors for visual confirmation of desired colonies are limited. Recently, we reported that TED (translational enhancement by a Dictyostelium gene sequence) boosted protein expression even without an expression inducer in Escherichia coli. Here, we demonstrate a generally applicable molecular tool using the expression of green fluorescent protein (GFP) enhanced by TED. By inserting a module related to TED into the cloning site in advance, we effectively screened E. coli colonies harboring the desired plasmid functions in a prokaryote (Magnetospirillum gryphiswaldense) or eukaryote (Dictyostelium discoideum). Thus, our system represents a user-friendly technique for cloning.

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Kondo, T., Yumura, S.. 2019-10-28. An improved molecular tool for screening bacterial colonies using GFP expression enhanced by a Dictyostelium sequence. https://doi.org/10.1101/821157

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