Fluorescence labeling and characterization of Lactiplantibacillus plantarum WLPL04
Lactiplantibacillus plantarum WLPL04,originated from healthy breast milk,can colonize the intestine.This arti-cle aimed to fluorescently label L.plantarum WLPL04,providing important experimental materials for studying its coloniza-tion,translocation,and distribution in vivo.In this study,the mCherry red fluorescent protein gene containing three different promoters(PX,P23 and Pldh)was used as the marker gene,and the recombinant plasmids pMG36 e-Px-mCherry,pMG36e-P23-mCherry and pMG36e-Pldh-mCherry were constructed.They were transformed into Escherichia coli and verified by PCR,plas-mid DNA digestion,colony color observation and laser confocal microscope observation.After successful validation and construc-tion,the three recombinant plasmids were further introduced into L.plantarum WLPL04.The results showed that L.plantarum WLPL04,which was introduced with plasmids pMG36e-Px-mCherry and pMG36e-P23-mCherry,showed significant red fluores-cence under laser confocal microscopy,while those carrying the pMG36e-Pldh-mCherry plasmid displayed weaker red fluores-cence.In addition,the effects of different recombinant plasmids on the growth and morphology of L.plantarum WLPL04 were observed and analyzed through growth curves and scanning electron microscopy.The results showed that the three plasmids had no significant effect on the growth and cell morphology of L.plantarum WLPL04.The above results indicated that L.plantarum WLPL04 has been successfully labeled with the red fluorescent protein mCherry,providing important experimental materials for subsequent research on its colonization,translocation,and distribution in vivo.
Lactiplantibacillus plantarum WLPL04mCherry fluorescent labelingpromoterexpression
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