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

During molecular cloning, screening 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 translational enhancement by a gene sequenc...

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Veröffentlicht in:BioTechniques 2020-02, Vol.68 (2), p.91-95
Hauptverfasser: Kondo, Tomo, Yumura, Shigehiko
Format: Artikel
Sprache:eng
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Zusammenfassung:During molecular cloning, screening 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 translational enhancement by a gene sequence (TED) boosted protein expression even without an expression inducer in . Here, we demonstrate a generally applicable molecular tool using the expression of green fluorescent protein enhanced by TED. By inserting a module related to TED into the cloning site in advance, we effectively screened colonies harboring the desired plasmid functions in a prokaryote ( ) or eukaryote ( ). Thus, our system represents a user-friendly technique for cloning.
ISSN:0736-6205
1940-9818
DOI:10.2144/btn-2019-0127