• Clinical and Experimental Pediatrics
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• 제58권1호
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• pp.20-27
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• 2015

Purpose: We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. Methods: Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. Results: Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. Conclusion: We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.226.2-226.2
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• 2003

The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically linked to the Itxa2b gene, which was then cloned into the pMAL -p2E expression vector. The chimaeric construction of pMALfimH/Itxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.309.2-309.2
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• 2002

The FimH subunit of type l-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically coupled to the ctxa2b gene, which was then cloned into pMAL -p2E expression vector. The chimaeric construction of pMALfimH/ctxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

• 한국진공학회:학술대회논문집
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• 한국진공학회 1997년도 제12회 학술발표회 논문개요집
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• pp.181-181
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• 1997

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.552-559
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• 2003

The FimH subunit of type 1-fimbriated Escherichiu coli (E. coli) has been determined as a major cause for urinary tract infections. Thus, to produce a possible vaccine antigen against urinary tract infections, the fimIH gene was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. A fusion protein, based on fusing adhesin to the cholera toxin subunit A2B (CTXA2B), was induced with 0.01 mM isopropyl-${\beta}-D-thiogalactoside$ (IPTG) for 4 h at $37^{\circ}C$ to yield a soluble fusion protein. The fusion protein was then purified by affinity chromatography. The expressed fusion protein was confirmed by SDS-PAGE and Western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was also analyzed. The orderly-assembled fusion protein was confirmed by a modified $G_{Ml}-ganglioside$ ELISA, using antibodies to adhesin. The results indicated that the purified fusion protein was an adhesin/CTXA2B protein containing E. coli adhesin and the $G_{Ml}-ganglioside$ binding activity of CTXB. Accordingly, this adhesin/CTXA2B protein may be a potential antigen for oral immunization against uropathogenic E. coli.

• Journal of Astronomy and Space Sciences
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• 제21권4호
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• pp.399-416
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• 2004

국내 최초의 과학위성인 과학기술위성 1호(STSAT-1)의 주 탑재체 원자외선분광기(FIMS; Far-utraviolet IMaging Spectrograph)는 2003년 9월 발사된 이후로, 다양한 관측을 수행하고 있으나 위성체에서 제공하는 자세정보의 시각오차의 문제점이 발견되어 시각오차를 보정하기 위한 연구가 수행되었다. 시각오차의 보정 후에 잘 알려진 천체의 관측결과를 이용하여 FIMS와 위성체 좌표계 사이의 기계적인 정렬오차에 의해 발행하는 FIMS의 시선방향과 위성체에서 제공하는 자세의 차이를 단파장대와 장파장대에 대하여 각각 계산하였다. 또한, 자세정보의 월별 상태 및 정밀도를 조사하였다. FIMS의 관측방법의 특성상 FIMS로부터 얻어진 영상은 위치별로 서로 다른 노출시간을 갖는다. 이러한 노출시간을 보정하는 방법을 기술하였다. 이 연구의 결과는 FIMS 자료분석을 위해 반드시 필요한 내용이며 추후 좀 더 정밀한 자세정보 보정을 위한 참고 자료로 활용될 것이다.

• Pediatric Infection and Vaccine
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• 제22권3호
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• pp.194-200
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• 2015

목적: 요로감염은 소아에서 흔한 세균 감염이며, Escherichia coli가 주요 원인균이다. 본 연구는 우리나라에서 소아 요로감염을 일으키는 E. coli의 계통 분류와 독성인자를 분석하고자 하였다. 방법: 2010년 10월부터 2013년 4월까지 요로감염으로 입원한 33명의 소아 환자로부터 검출된 E. coli균주를 대상으로 하였다. 중합효소연쇄반응을 통해 E. coli의 계통 분류 및 5가지 독성인자(fimH, sfa, papA, hylA, and cnf1)를 조사하였다. E. coli의 분자유전학적 특징을 환자의 임상적 진단과 동반된 방광요관 역류에 따라 분석하였다. 결과: 대부분의 요로병원성 E. coli 는 계통 분류에서 B2군(84.8%)에 속했으며, 나머지는 모두 D군(15.2%)에 해당되었다. 독성인자는 fimH (100%), sfa (100%), hylA (63.6%), cnfI (63.6%), 그리고 papA (36.4%)의 분포를 보였다. 임상 진단에 따른 계통 분류에서 급성 신우신염의 경우 B2군이 92.3%, D군이 7.7%를 나타냈으며, 방광염에서는 B2군에서 57.1%, D2군은 42.9%였다. 독성인자는 양 군에서 비슷하게 분포하였다. 급성 신우신염에서 방광요관 역류의 유무에 따른 계통 분류의 분포에는 차이가 없었으나, 독성인자의 경우 papA 유전자가 방광요관 역류가 동반되지 않은 군에서보다 방광요관 역류 군에서 적게 나타났다(43.8% vs. 20.0%, P=0.399). 결론: 본 연구는 국내 소아 요로감염의 원인 E. coli 균주의 분자유전학적 역학 자료를 제시하였으며, 이 결과는 향후 소아 요로감염의 발생 기전을 이해하는 데 기초가 될 것으로 생각된다.

• Journal of Animal Science and Technology
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• 제46권5호
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• pp.841-848
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• 2004

살균된 우유에 오염된 Salmonella의 신속한 검출 방법을 공중 보건의 보호를 위하여 중요하다. RT-PCR 방법은 생균과 사균을 분별하여 검출할 수 있는 분자유전학적 방법이다. 본 연구의 RT-PCR 방법은 Salmonella의 type 1 fImbriae의 단량체 단백질을 암호화하는 fimA 유전자의 mRNA를 주형으로 하여 DNA를 증폭하는 방법으로서 활력이 있는 Salmonella를 검출할수 있기 위하여 개발되었다. RNA 시료를 RQRNase-free DNase로 처리하여 오염된 DNA를 파괴하여 RT-PCR 반응에서 주형으로서 DNA가 합성되는 것을 방지할 수 있었다. 이 RT-PCR 방법은 Salmonella 7균주를 검출하였으나 Escherichia coli, Shigella sonnei, Citrobacter freundii, 및 Klebsiella pneumoniae 는 검출하지 않았다. 우유 내에서 열처리된 $10^7/ml과 10^6/ml$ 세균수의 Salmonella는 검출되 었으나 $10^5{\sim}100/ml$는 검출되지 않았다. RT-PCR를 검출할 수 있는 활력이 있는 Salmonella의 최소 세균수는 100/ml이었다.

• Biomolecules & Therapeutics
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• 제12권3호
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• pp.179-184
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• 2004

The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause for urinary tract infections. In our previous study, the Adhesin/CTXA2B was expressed as soluble recombinant chimaeric protein derived from the uropathogenic Escherichia coli adhesin genetically coupled to cholera toxin A2B (CTXA2B) subunit in Escherichia coli. Since it is very important to optimize IPTG concentration and culture temperature to maximize cell growth and productivity, These optimal culture factors were determined to increase the productivity of the expressed Adhesin/CTXA2B chimaeric protein in Escherichia coli TB1 carrying pMALfimH/ctxa2b. Our data demonstrate that optimal concentration of IPTG for increased production of chimaeric protein was 0.5 mM. Additionally, culture time was 10 hours and temperature, 37${\circ}C$.

• BMB Reports
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• 제37권3호
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• pp.314-319
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• 2004

Flounder (Paralichthys olivaceus) Immunoglobulins (Igs) were purified from the serum of mouse IgG-immunized flounder by using affinity chromatography. Under denaturing conditions in SDS-PAGE, the flounder Igs appeared to be composed of 2 heavy (H) chains (72 and 77 kDa) and two light (L) chains (26 and 28 kDa). Monoclonal antibodies (MAbs) were produced by the fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder Igs. In a Western blot analysis, the produced MAbs, FIM511, FIM519, and FIM562 recognized both the 72 and 77 kDa H chains, 26 kDa, and 28 kDa L chain, respectively. Mouse antiserum against flounder Igs reacted more strongly with the L chain of 28 kDa than with 26 kDa, suggesting that the 28 kDa molecule is more immunogenic than the 26 kDa L chain molecule. In a FACS analysis, the ratios of the Ig+ cell population in the flounder head kidney and spleen cells were 49% and 24%, respectively. Unexpectedly, however, the ratios of the Ig+ B-like cell population in the flounder were not significantly augmented, even after the immunization of an immunogenic antigen. This suggests that the humoral immune response in fish could be considerably different from that in mammals. The produced MAbs in this study would be useful in characterizing flounder Ig+ B-like cells and in developing flounder Ig detecting an immunoassay system.