수해양성 병원성 미생물로 알려진 Vibrio anguillarum으로부터 mannose-1-phosphate를 mannose-6-phosphate, glucose-1-phosphate를 glucose-6-phosphate로 가역적으로 변환시키는 phosphomannomutase/phosphoglucomutase (pmm/pgm)의 ...
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https://www.riss.kr/link?id=A102118263
2016
Korean
KCI등재,SCOPUS
학술저널
355-362(8쪽)
0
0
상세조회0
다운로드국문 초록 (Abstract)
수해양성 병원성 미생물로 알려진 Vibrio anguillarum으로부터 mannose-1-phosphate를 mannose-6-phosphate, glucose-1-phosphate를 glucose-6-phosphate로 가역적으로 변환시키는 phosphomannomutase/phosphoglucomutase (pmm/pgm)의 ...
수해양성 병원성 미생물로 알려진 Vibrio anguillarum으로부터 mannose-1-phosphate를 mannose-6-phosphate, glucose-1-phosphate를 glucose-6-phosphate로 가역적으로 변환시키는 phosphomannomutase/phosphoglucomutase (pmm/pgm)의 유전자를 sequencing하여 1338 bp의 open reading frame (ORF)을 밝혔다. 이는 446개의 아미노산을 포함하며 47,625 Da을 가지고 있다. 보고된 다른 Vibrio sp. 의 pmm/pgm 유전자와 상동성을 비교하였을 때 V. mimicus V. vulnificus, V. splendidus, V. harveyi와 92.3%, 91.4%, 89.9%, 89.9%에 해당하는 상동성을 지니고 있었다. 증폭된 목적 유전자를 pET-28a(+) vector에 연결하여 대장균에서 단백질의 대량발현을 유도하였으며 이는 주로 soluble한 상태로 나왔다. Soluble fraction을 Ni-NTA column chromatography로 정제하여 약 50 kDa의 단백질을 얻었고 이는 주로 mannose-1-phosphate를 이용하는 효소로 확인되었으며 Mg2+ 이온이 존재할 때 효소의 활성이 나타나는 것을 확인할 수 있었다. 본 연구의 유전자는 낮은 온도의 stress하에서 발현이 증가됨을 Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR)을 통해 확인하였고, 상동성 재조합 (homologous recombination)에 의한 돌연변이 균주 제작을 통해 PMM/PGM protein과 lipopolysaccharide (LPS)의 생합성과의 관계를 규명하였다. V. anguillarum wild type과 mutant로부터 LPS를 분리하였고 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE)후 silver staining을 통해 LPS의 high molecular weight (HMW) 부분인 O-antigen에서의 변화를 확인하였다. 또한 V. anguillarum wild type과 mutant의 growth와 viability를 확인한 결과 mutant가 wild type보다 정지기까지 더 낮은 생육을 보였으며 viability가 감소함을 확인하였다. 본 연구를 통하여 V. anguillarum의 pmm/pgm 유전자가 미생물의 생육과 LPS 생합성에 관여하고 있음을 알 수있었다.
다국어 초록 (Multilingual Abstract)
The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree ...
The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose 1 phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.
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1 Actis LA, "vol. 3: viral, bacterial, and fungal infections, Vibriosis" 570-605, 2011
2 Larsen, JL, "Vibrio anguillarum serovars associated with vibriosis in fish" 17 : 259-267, 1994
3 Frans I, "Vibrio anguillarum as a fish pathogen: virulence factors, diagnosis and prevention" 34 : 643-661, 2011
4 Banemann A, "The lipopolysaccharide of Bordetella bronchiseptica acts as a protective shield against antimicrobial peptides" 66 : 5607-5612, 1998
5 Preston A, "The lipoligosaccharides of pathogenic Gram-negative bacteria" 22 : 139-180, 1996
6 Ray WJ, "The enzymes, vol. 6" Academic Press 407-477, 1972
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Lipopolysaccharide로 유도된 RAW 264.7 세포와 마우스모델에 대한 진두발 에탄올 추출물의 항염증 효과
국내외 시판 농산물 중간소재의 총페놀, 총플라보노이드, 총안토시아닌 함량 및 항산화 활성
In vitro 실험을 통한 벼세포 추출물의 창상 치유 효능 평가
학술지 이력
연월일 | 이력구분 | 이력상세 | 등재구분 |
---|---|---|---|
2023 | 평가예정 | 해외DB학술지평가 신청대상 (해외등재 학술지 평가) | |
2020-01-01 | 평가 | 등재학술지 유지 (해외등재 학술지 평가) | |
2015-09-23 | 학술지명변경 | 외국어명 : Korean Journal of Microbiology and Biotechnology -> Microbiology and Biotechnology Letters | |
2010-01-01 | 평가 | 등재 1차 FAIL (등재유지) | |
2008-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2006-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2004-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2001-01-01 | 평가 | 등재학술지 선정 (등재후보2차) | |
1998-07-01 | 평가 | 등재후보학술지 선정 (신규평가) |
학술지 인용정보
기준연도 | WOS-KCI 통합IF(2년) | KCIF(2년) | KCIF(3년) |
---|---|---|---|
2016 | 0.6 | 0.6 | 0.65 |
KCIF(4년) | KCIF(5년) | 중심성지수(3년) | 즉시성지수 |
0.53 | 0.55 | 0.977 | 0.18 |