title: Intracellular imaging of Qdots-labeled DNA in cyanobacteria.
abstract: In this contribution, they have attempted to develop a labeling technique for in vivo imaging of functionally active plasmid DNA in cyanobacterial cells through its decoration with semiconductor quantum dots (Qdots) as fluorescent nanoprobes. For that purpose biotinylated plasmid slr2060 DNA was conjugated with Qdots-streptavidine. The intact DNA was visualized in a single green color by light microscopy. These Qdots-DNA conjugates were capable of expressing the acyltransferase enzyme. Qdots-DNA conjugates and confocal microscope imaging technique were adopted to visualize the gene transport across the membrane of the live cyanobacteria cell in real time. Long-term kinetic study enabled to reveal the steps of extracellular and intracellular microenvironment for plasmid transportation into the live cell. To confirm these processes a confocal microscope and indicator plate assay test were applied in tandem. In this contribution, Qdots-labeled plasmid DNA was utilized for the first time for long-term intracellular imaging studies in cyanobacteria species PCC6803. The results showed that the Qdots-labeled plasmid DNA detection could be used as a powerful labeling technique for visualization of exogenous DNA entry and tracking into living cells by confocal microscopy.
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-Ohkubo 255, Sakura - Ku, Saitama, 338-8570, Japan.|Laboratory of Engineering NanoBiotechnology, Department of Engineering geoecology, University of Mining and Geology "St. Ivan Rilski", Sofia, Bulgaria.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Loukanov
authors.affiliation: Laboratory of Engineering NanoBiotechnology, Department of Engineering geoecology, University of Mining and Geology "St. Ivan Rilski", Sofia, Bulgaria.
authors.forename: Veselin
authors.initials: V
authors.lastname: Zhelyazkov
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-Ohkubo 255, Sakura - Ku, Saitama, 338-8570, Japan.
authors.forename: Yukako
authors.initials: Y
authors.lastname: Hihara
authors.affiliation: Graduate School of Science and Engineering, Saitama University, Shimo-Ohkubo 255, Sakura - Ku, Saitama, 338-8570, Japan.
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Nakabayashi
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D000458:Cyanobacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry*; D018613:Microscopy, Confocal; D045663:Quantum Dots / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial
keywords: Qdots-labeled plasmid DNA; cyanobacteria Synechocystis sp. PCC6803; in vivo gene tracking; slr2060 acyltransferase