Journal of Microbiology

Journal Article

Colonization study of gfp-tagged Achromobacter marplatensis strain in sugar beet: YingWu Shi , Chun Li Li , HongMei Yang , Tao Zhang , Yan Gao , Min Chu , Jun Zeng , Qing Lin , OuTiKu Er , YuGuo Li , Xiangdong Huo , Kai Lou; J. Microbiol. 2017;55(4):267-272. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6371-1

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This study details the introduction of a gfp marker into an endophytic bacterial strain (Achromobacter marplatensis strain 17, isolated from sugar beet) to monitor its coloniza-tion of sugar beet (Beta. vulgaris L.). Stability of the plasmid encoding the gfp was confirmed in vitro for at least 72 h of bacterial growth and after the colonization of tissues, under nonselective conditions. The colonization was observed us-ing fluorescence microscopy and enumeration of culturable endophytes in inoculated sugar beet plants that grew for 10 or 20 days. gfp-Expressing strains were re-isolated from the inner tissues of surface-sterilized roots and stems of inocu-lated plants, and the survival of the Achromobacter marpla-tensis 17:gfp strain in plants 20 days after inoculation, even in the absence of selective pressure, suggests that it is good colonizer. These results also suggest that this strain could be a useful tool for the delivery of enzymes or other proteins into plants. In addition, the study highlights that sugar beet plants can be used effectively for detailed in vitro studies on the interactions between A. marplatensis strain 17 and its host, particularly if a gfp-tagged strain of the pathogen is used.

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Research Support, Non-U.S. Gov'ts

NOTE] GFP-Expressing Influenza A Virus for Evaluation of the Efficacy of Antiviral Agents: Jin Il Kim , Sehee Park , Ilseob Lee , Sangmoo Lee , Saem Shin , Yongkwan Won , Min-Woong Hwang , Joon-Yong Bae , Jun Heo , Hye-Eun Hyun , Hyejin Jun , Soon Sung Lim , Man-Seong Park; J. Microbiol. 2012;50(2):359-362. Published online April 27, 2012; DOI: https://doi.org/10.1007/s12275-012-2163-9

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Abstract: To address its value as a screening tool in the development of antiviral drugs, a recombinant influenza virus expressing green fluorescent protein (rPR8-GFP virus) was investigated in vitro and in vivo. The inhibition of viral growth by a neuraminidase inhibitor in the cells or lower respiratory tracts of mice could be visualized by the level of fluorescence. In addition, the rPR8-GFP virus exhibited high pathogenicity in mice. Taken together, these results suggest that the rPR8-GFP virus can be a useful tool for the rapid identification of antiviral drugs active against influenza viruses.

Translocation of Green Fluorescent Protein to Cyanobacterial Periplasm Using Ice Nucleation Protein: Wipa Chungjatupornchai , Sirirat Fa-aroonsawat; J. Microbiol. 2009;47(2):187-192. Published online May 2, 2009; DOI: https://doi.org/10.1007/s12275-008-0188-x

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Abstract

The translocation of proteins to cyanobacterial cell envelope is made complex by the presence of a highly differentiated membrane system. To investigate the protein translocation in cyanobacterium Synechococcus PCC 7942 using the truncated ice nucleation protein (InpNC) from Pseudomonas syringae KCTC 1832, the green fluorescent protein (GFP) was fused in frame to the carboxyl-terminus of InpNC. The fluorescence of GFP was found almost entirely as a halo in the outer regions of cells which appeared to correspond to the periplasm as demonstrated by confocal laser scanning microscopy, however, GFP was not displayed on the outermost cell surface. Western blotting analysis revealed that InpNC-GFP fusion protein was partially degraded. The N-terminal domain of InpNC may be susceptible to protease attack; the remaining C-terminal domain conjugated with GFP lost the ability to direct translocation across outer membrane and to act as a surface display motif. The fluorescence intensity of cells with periplasmic GFP was approximately 6-fold lower than that of cells with cytoplasmic GFP. The successful translocation of the active GFP to the periplasm may provide a potential means to study the property of cyanobacterial periplasmic substances in response to environmental changes in a non-invasive manner.

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Stefano Ferri, Mayumi Nakamura, Akiko Ito, Mitsuharu Nakajima, Koichi Abe, Katsuhiro Kojima, Koji Sode
Algal Research.2015; 12: 337. CrossRef
Comparative Mechanisms of Protein Transduction Mediated by Cell-Penetrating Peptides in Prokaryotes
Betty Revon Liu, Yue-Wern Huang, Robert S. Aronstam, Han-Jung Lee
The Journal of Membrane Biology.2015; 248(2): 355. CrossRef
Construction of a cell-surface display system based on the N-terminal domain of ice nucleation protein and its application in identification of mycoplasma adhesion proteins
S. Bao, S. Yu, X. Guo, F. Zhang, Y. Sun, L. Tan, Y. Duan, F. Lu, X. Qiu, C. Ding
Journal of Applied Microbiology.2015; 119(1): 236. CrossRef
The rrnA promoter as a tool for the improved expression of heterologous genes in cyanobacteria
Wipa Chungjatupornchai, Sirirat Fa-aroonsawat
Microbiological Research.2014; 169(5-6): 361. CrossRef
Arabinogalactan Proteins Occur in the Free-Living Cyanobacterium Genus Nostoc and in Plant–Nostoc Symbioses
Owen Jackson, Oliver Taylor, David G. Adams, J. Paul Knox
Molecular Plant-Microbe Interactions®.2012; 25(10): 1338. CrossRef
Translocation of green fluorescent protein by comparative analysis with multiple signal peptides
Elisabeth Linton, Marie K. Walsh, Ronald C. Sims, Charles D. Miller
Biotechnology Journal.2012; 7(5): 667. CrossRef
Display of Organophosphorus Hydrolase on the Cyanobacterial Cell Surface Using Synechococcus Outer Membrane Protein A as an Anchoring Motif
Wipa Chungjatupornchai, Attapon Kamlangdee, Sirirat Fa-aroonsawat
Applied Biochemistry and Biotechnology.2011; 164(7): 1048. CrossRef

Evaluation of Endophytic Colonization of Citrus sinensis and Catharanthus roseus Seedlings by Endophytic Bacteria: Paulo Teixeira Lacava , Welington Luiz Araujo , Joao Lucio Azevedo; J. Microbiol. 2007;45(1):11-14.; DOI: https://doi.org/2498 [pii]

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Abstract: Over the last few years, the endophytic bacterial community associated with citrus has been studied as an important component interacting with Xylella fastidiosa, the causal agent of citrus variegated chlorosis (CVC). This bacterium may also colonize some model plants, such as Catharanthus roseus and Nicotiana clevelandii. In the present study, we compared the endophytic colonization of Citrus sinensis and Catharanthus roseus using the endophytic bacteria Klebsiella pneumoniae. We chose an appropriate strain, K. pneumoniae 342 (Kp342), labeled with the GFP gene. This strain was inoculated onto seedlings of C. sinensis and C. roseus. The isolation frequency was determined one week after the inoculation and the endophytic colonization of K. pneumoniae was observed using fluorescence microscopy. Although the endophytic bacterium was more frequently isolated from C. roseus than from C. sinensis, the colonization profiles for both host plants were similar, suggesting that C. roseus could be used as a model plant to study the interaction between endophytic bacteria and X. fastidiosa.

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