Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Bifidobacterium are recognized as crucial beneficial gut microorganisms.
• Understanding Bifidobacterium roles requires effective methods for their study.
• Current visualization techniques for Bifidobacterium may be limited.

Purpose of the Study:

• To develop a novel bioluminescent plasmid vector for Bifidobacterium.
• To enable efficient visualization of Bifidobacterium strains.
• To facilitate studies of Bifidobacterium in vitro, in situ, and in vivo.

Main Methods:

• A plasmid replicon from Bifidobacterium longum FI10564 was amplified and cloned.
• The replicon was subcloned into a pTG262 vector containing a luciferase gene (luc+).
• The final recombinant plasmid (pTG262::pFI2576 rep (luc+)) was introduced into Bifidobacterium catenulatum.

Main Results:

• The recombinant Bifidobacterium catenulatum strain exhibited significant luminescence.
• A relative luminescence of 3,024 units was recorded at OD600 = 0.352.
• The developed plasmid construct successfully labeled the Bifidobacterium strain.

Conclusions:

• A functional bioluminescent plasmid vector for Bifidobacterium has been successfully constructed.
• This recombinant plasmid enables broad application for labeling Bifidobacterium species.
• The tool will aid in the visualization and study of Bifidobacterium in diverse settings.