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Bacterial Surface Display of $GFP_{UV}$ on Bacillus subtilis Spores  

Kim, Jung-Hyung (Department of Chemical Engineering, Dong-A University)
Roh, Chang-Hyun (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University)
Lee, Chang-Won (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University)
Kyung, Do-Hyun (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University)
Choi, Soo-Keun (Genofocus Inc.)
Jung, Heung-Chae (Genofocus Inc.)
Pan, Jae-Gu (Genofocus Inc.)
Kim, Byung-Gee (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.17, no.4, 2007 , pp. 677-680 More about this Journal
Abstract
To analyze a cotG-based Bacillus subtilis spore display system directly, $GFP_{UV}$ was expressed on the surface of Bacillus subtilis spores. When $GFP_{UV}$ was fused to the C-terminal of the cotG structural gene and expressed, the existence of a $CotG-GFP_{UV}$ fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This $GFP_{UV}$ displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.
Keywords
Bacillus subtilis; cotG; $GFP_{UV}$; spore; surface display;
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Times Cited By KSCI : 3  (Citation Analysis)
Times Cited By Web Of Science : 7  (Related Records In Web of Science)
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1 Crameri, A., E. A. Whitehorn, E. Tate, and W. P. Stemmer. 1996. Improved green fluorescent protein by molecular evolution using DNA shuffiing. Nat. Biotechnol. 14: 315-319   DOI   ScienceOn
2 Jung, H. C., J. M. Lebeault, and J. G. Pan. 1998. Surface display of Zymomonas mobilis levansucrase by using the ice-nucleation protein of Pseudomonas syringae. Nat. Biotechnol. 16: 576-580   DOI   ScienceOn
3 Kawamura, F. and R. H. Doi. 1984. Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. J. Bacteriol. 160: 442-444
4 Lee, Y., E. Ahn, S. Park, E. L. Masden, C. O. Jeon, and W. Park. 2006. Construction of a reporter strain Pseudomonas putida for the detection of oxidative stress caused by environmental pollutants. J. Microbiol. Biotechnol. 16: 386-390   과학기술학회마을
5 Kim, J. H., C. S. Lee, and B. G. Kim. 2005. Spore-displayed streptavidin: A live diagnostic tool in biotechnology. Biochem. Biophys. Res. Commun. 331: 210-214   DOI   ScienceOn
6 Sousa, C., A. Cebolla, and V. de Lorenzo. 1996. Enhanced metalloadsorption of bacterial cells displaying poly-His peptides. Nat. Biotechnol. 14: 1017-1020   DOI   ScienceOn
7 Shi, H. and W. Wen Su. 2001. Display of green fluorescent protein on Escherichia coli cell surface. Enzyme Microb. Technol. 28: 25-34   DOI   ScienceOn
8 Choi, J. H., J. I. Choi, and S. Y. Lee. 2005. Display of proteins on the surface of Escherichia coli by C-terminal deletion fusion to the Salmonella typhimurium OmpC. J. Microbiol. Biotechnol. 15: 141-146   과학기술학회마을
9 Kim, H., M. Hahn, P. Grabowski, D. C. McPherson, M. M. Otte, R. Wang, C. C. Ferguson, P. Eichenberger, and A. Driks. 2006. The Bacillus subtilis spore coat protein interaction network. Mol. Microbiol. 59: 487-502   DOI   ScienceOn
10 Ye, K., S. Shibasaki, M. Ueda, T. Murai, N. Kamasawa, M. Osumi, K. Shimizu, and A. Tanaka. 2000. Construction of an engineered yeast with glucose-inducible emission of green fluorescence from the cell surface. Appl. Microbiol. Biotechnol. 54: 90-96   DOI   ScienceOn
11 Richins, R. D., I. Kaneva, A. Mulchandani, and W. Chen. 1997. Biodegradation of organophosphorus pesticides by surface-expressed organophosphorus hydrolase. Nat. Biotechnol. 15: 984-987   DOI   ScienceOn
12 Georgiou, G., C. Stathopoulos, P. S. Daugherty, A. R. Nayak, B. L. Iverson, and R. Curtiss 3rd. 1997. Display of heterologous proteins on the surface of microorganisms: From the screening of combinatorial libraries to live recombinant vaccines. Nat. Biotechnol. 15: 29-34   DOI   ScienceOn
13 Li, L., D. G. Kang, and H. J. Chao 2004. Functional display of foreign protein on surface of Escherichia coli using, N-terminal domain of ice nucleation protein. Biotechnol. Bioeng. 85: 214-221   DOI   ScienceOn
14 Kim, J., S. Kim, C. O. Jeon, J. Yun, H. S. Lee, and H. S. Ro. 2006. Screening of yeast diauxic promoters for production of foreign proteins. J. Microbiol. Biotechnol. 16: 1459-1463   과학기술학회마을
15 Boder, E. T. and K. D. Wittrup. 1997. Yeast surface display for screening combinatorial polypeptide libraries. Nat. Biotechnol. 15: 553-557   DOI   ScienceOn
16 Harwood, C. and S. Cutting. 1990. Molecular Biological Methods for Bacillu. John Wiley & Sons, Chichester
17 Chapple, S. D. and I. M. Jones. 2002. Non-polar distribution of green fluorescent protein on the surface of Autographa californica nucleopolyhedrovirus using a heterologous membrane anchor. J. Biotechnol. 95: 269-275   DOI   ScienceOn
18 Sacco, M., E. Ricca, R. Losick, and S. Cutting. 1995. An additional GerE-controlled gene encoding an abundant spore coat protein from Bacillus subtilis. J. Bacteriol. 177: 372-377   DOI
19 Goyal, A. and J. K. Batra. 2000. Inclusion of a furin-sensitive spacer enhances the cytotoxicity of ribotoxin restrictocin containing recombinant single-chain immunotoxins. Biochem. J. 345 Pt 2: 247-254   DOI   ScienceOn
20 Ojala, K., J. Koski, W. Ernst, R. Grabherr, I. Jones, and C. Oker-Blom. 2004. Improved display of synthetic IgG-binding domains on the baculovirus surface. Technol. Cancer Res. Treat. 3: 77-84   DOI
21 Shibasaki, S., A. Tanaka, and M. Ueda. 2003. Development of combinatorial bioengineering using yeast cell surface display -- order-made design of cell and protein for bio-monitoring. Biosens. Bioelectron. 19: 123-130   DOI   ScienceOn