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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제26권 제1호
- 발행연도
- 2016.1
- 수록면
- 160 - 170 (11page)
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초록· 키워드
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The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.
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참고문헌 (42)
참고문헌 신청
Borisjuk NV / 1999 / Production of recombinant proteins in plant root exudates / Nat. Biotechnol 17:466~469
Borysowski J / 2006 / Bacteriophage endolysins as a novel class of antibacterial agent / Exp. Biol. Med 231:366~377
Chapman S / 1992 / Potato virus X as a vector for gene expression in plants / Plant J 2:549~557
Company N / 2014 / The production of recombinant cationic α-helical antimicrobial peptides in plant cells induces the formation of protein bodies derived from the endoplasmic reticulum / Plant Biotechnol. J 12:81~92
Daniell H / 2001 / Expression of the native cholera B toxin subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts / J. Mol. Biol 311:1001~1009
Borysowski J / 2006 / Bacteriophage endolysins as a novel class of antibacterial agent / Exp. Biol. Med 231:366~377
Chapman S / 1992 / Potato virus X as a vector for gene expression in plants / Plant J 2:549~557
Company N / 2014 / The production of recombinant cationic α-helical antimicrobial peptides in plant cells induces the formation of protein bodies derived from the endoplasmic reticulum / Plant Biotechnol. J 12:81~92
Daniell H / 2001 / Expression of the native cholera B toxin subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts / J. Mol. Biol 311:1001~1009
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