• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.172-175
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• 2002

The complete nucleotide sequence of the nahC gene from Pseudomonas fluorescens, the structural gene for 1,2-dihydroxynaphthalene (1,2-DHN) dioxygenase, was determined. The 1,2-DHN dioxygenase is an extradiol ring-cleavage enzyme that cleaves the first ring of 1,2-dihydroxynaphthalene. The amino acid sequence of the dioxygenase deduced from the nucleotide sequence suggested that the holoenzyme consists of eight identical subunits with a molecular weight of approximately 34,200. The amino acid sequence of 1,2-DHN dioxygenase showed more than $90\%$ homology with those of the dioxygenases of other Pseudomonas strains. However, sequence similarity with those of the Sphingomonas species was less than $60\%$. The nahC gene of P. fluorescens was moderately expressed in E. coli NM522, as determined by enzymatic activity.

• 한국미생물·생명공학회지
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• 제17권1호
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• pp.51-55
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• 1989

TOL 플라스미드와 NAH 플라스미드는 n-알칸을 자화하는 P. putida KCTC 2405에 접합에 의해 각각의 이동은 가능하나 두 플라스미드는 불화합성에 기인하여 본 균주내에 공존할 수 없었다. TOL plasmid에서 불화합성 체계는 남겨두고 tol 유전자만 이 CAM plasmid내로 transposition 되어 형성된 CAM::TOL* 플라스미드는 NAH 플라스미드와 P. putida KCTC 2405에서 공존할 수 있어 m-toluate, naphthalene, camphor 및 n-alkane(C8-C24)를 분해할 수 있는 P. putida 3SK 균주를 육종하였다. CAM::TOL* 플라스미드는 선택성 배지에서 안정하였으나 비선택성 배지에서는 불안정하였다.

• Journal of Ginseng Research
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• 제29권4호
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• pp.167-175
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• 2005

The $Ca^{2+}-activated$ chloride channel (CLCA) was activated by ginseng total saponin (GTS) in Xenopus oocytes. The reverse transcription PCR (RT-PCR) method was performed with gene specific primers on oocytes. The gene specific primers were deduced from spleen cDNA in expressed sequence tags (EST) database showing high homology to the mouse CLCA. Full length of cDNA sequence was completed by linkage of several 5' and 3'-half cDNA fragments have been sequenced. We named the full cDNA to oCLCA transiently. The oCLCA gene encodes a protein of 911 amino acids with $48.9\%$ identity overall to that of mouse CLCA (mCLCA4). A predicted oCLCA amino acids sequence shows the molecular weight of 108 kDa and has four or more transmembrane domains, and also the one hydrophobic C­terminal domain. oCLCA gene was expressed ubiquitously in various tissues included oocytes, also interfered in oocytes by siRNA for oCLCA. Here, we suggest that oCLCA is a endogenous chloride channel gene in oocytes. We are studying for the identification of oCLCA gene and further physiological research.

• Journal of Microbiology
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• 제44권5호
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• pp.473-479
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• 2006

A well characterized naphthalene-degrading strain, Pseudomonas putida PpG7 was observed to utilize limonin, a highly-oxygenated triterpenoid compound as a sole source of carbon and energy. Limonin concentrations evidenced a 64% reduction over 48 h of growth in batch cultures. Attempts were made to acquire a plasmid-less derivative via various methods (viz. Ethidium Bromide, SDS, elevated temperature & mitomycin C), among which the method involving mitomycin C ($20{\mu}g/ml$) proved successful. Concomitant with the loss of plasmid in P. putida PpG7 strain, the cured derivative was identified as a $lim^-$ phenotype. The $lim^+$ phenotype could be conjugally transferred to the cured derivative. Based on the results of curing with mitomycin C, conjugation studies and presence of ndo gene encoding naphthalene 1,2 dioxygenase, it was demonstrated that genes for the limonin utilization were encoded on an 83 kb indigenous transmissible Inc. P9 NAH plasmid in Pseudomonas putida PpG7 strain.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.26-26
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• 2015

Phytophthora infestans is the causal agent of potato and tomato late blight, one of the most devastating plant diseases. P. infestans secretes effector proteins that are both modulators and targets of host plant immunity. Among these are the so-called RXLR effectors that function inside plant cells and are characterized by a conserved motif following the N-terminal signal peptide. In contrast, the effector activity is encoded by the C terminal region that follows the RXLR domain. Recently, I performed in planta functional profiling of different RXLR effector alleles. These genes were amplified from a variety of P. infestans isolates and cloned into a Potato virus X (PVX) vector for transient in planta expression. I assayed for R-gene specific induction of hypersensitive cell death. The findings included the discovery of new effector with avirulence activity towards the Solanum bulbocastanum Rpi-blb2 resistance gene. The Rpi-blb2 encodes a protein with a putative CC-NBS-LRR (a coiled-coil-nucleotide binding site and leucine-rich repeat) motif that confers Phytophthora late blight disease resistance. We examined the components required for Rpi-blb2-mediated resistance to P. infestans in Nicotiana benthamiana. Virus-induced gene silencing was used to repress candidate genes in N. benthamiana and to assay against P. infestans infections. NbSGT1 was required for disease resistance to P. infestans and hypersensitive responses (HRs) triggered by co-expression of AVRblb2 and Rpi-blb2 in N. benthamiana. RAR1 and HSP90 did not affect disease resistance or HRs in Rpi-blb2-transgenic plants. To elucidate the role of salicylic acid (SA) in Rpi-blb2-mediated resistance, we analyzed the response of NahG-transgenic plants following P. infestans infection. The increased susceptibility of Rpi-blb2-transgenic plants in the NahG background correlated with reduced SA and SA glucoside levels. Furthermore, Rpi-blb2-mediated HR cell death was associated with $H_2O_2$, but not SA, accumulation. SA affects basal defense and Rpi-blb2-mediated resistance against P. infestans. These findings provide evidence about the roles of SGT1 and SA signaling in Rpi-blb2-mediated resistance against P. infestans.

• 미생물학회지
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• 제30권1호
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• pp.8-14
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• 1992

Pseudomonas sp. DJ77로부터 extradiol dioxygenase 유전자(phnE)를 클로닝하고 염기배열을 결정하였다. 921 bp의 open reading frame (ORF) 이 존재하였고 개시코돈 앞에서 Shine-Dalgarno sequence를 발견하였다. phnE 유전자에서 만들어지는 PhnE 단백질은 분자량이 34,449 Da 인데 SDS-polycrylamide gel 전기영동에 의해 측정된 분자량과 일치하였다. PhnE는 NahH, XylE, DmpB 등과 아미노산 배열의 상동성의 약 50% 였다. DJ77에는 bphC와 같은 3형의 extradiol dioxygenase 유전자는 발견할 수 없었다. DJ77 과 JM101(pPE17)은 catechol, 3-methylcatechol, 4-methylcatechol, 2, 3-dihydroxybiphenyl 등의 기질을 meta-cleavage 하여 노란색 화합물을 생성할 수 있었다.

• Macromolecular Research
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• 제12권6호
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• pp.573-580
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• 2004

To prepare chitosan-based polymeric amphiphiles that can form nanosized core-shell structures (nanopar­ticles) in aqueous milieu, chitosan oligosaccharides (COSs) were modified chemically with hydrophobic cholesterol groups. The physicochemical properties of the hydrophobized COSs (COSCs) were investigated by using dynamic light scattering and fluorescence spectroscopy. The feasibility of applying the COSCs to biomedical applications was investigated by introducing them into a gene delivery system. The COSCs formed nanosized self-aggregates in aqueous environments. Furthermore, the physicochemical properties of the COSC nanoparticles were closely related to the molecular weights of the COSs and the number of hydrophobic groups per COS chain. The critical aggregation concentration values decreased upon increasing the hydrophobicity of the COSCs. The COSCs effi­ciently condensed plasmid DNA into nanosized ion-complexes, in contrast to the effect of the unmodified COSs. An investigation of gene condensation, performed using a gel retardation assay, revealed that $COS6(M_n=6,040 Da)$ containing $5\%$ of cholesteryl chloroformate (COS6C5) formed a stable DNA complex at a COS6C5/DNA weight ratio of 2. In contrast, COS6, the unmodified COS, failed to form a stable COS/DNA complex even at an elevated weight ratio of 8. Furthermore, the COS6C5/DNA complex enhanced the in vitro transfection efficiency on Human embryonic kidney 293 cells by over 100 and 3 times those of COS6 and poly(L-lysine), respectively. Therefore, hydrophobized chitosan oligosaccharide can be considered as an efficient gene carrier for gene delivery systems.

• 대한전자공학회:학술대회논문집
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• 대한전자공학회 2003년도 하계종합학술대회 논문집 Ⅳ
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• pp.1743-1746
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• 2003

C. elegans often used to study of function of gene, but it is difficult for human observation to distinguish the mutants of C. elegans. To solve this problem, the system, which can be classified automatically using the computer vision, is studying now. In the previous works , they described the auto-tracking system and the egg-laying timing modeling, which are used to automated-classily system. In this paper, we use three kinds of features, which are related to movement , size and posture of the worm, and each feature is described mathematically and normalized. In experimental result, we validated the features for the hierarchical clustering, And we used the Calinski and Harabasz's method to find the appropriate cluster number.

• 미생물학회지
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• 제38권4호
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• pp.275-275
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• 2002

Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as carbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

• 한국통신학회논문지
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• 제28권7C호
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• pp.712-718
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• 2003

C.elegans 선충은 유전자 기능 연구에 주로 쓰이고 있으나, 변종들의 구분이 육안으로는 쉽지 않다. 이를 해결하기 위하여 컴퓨터 비젼을 이용하여 자동으로 분류할 수 있는 시스템이 연구 중이며, 이전 논문［1］에서 선충의 자동 분류 시스템에 사용될 영상의 전처리 과정에 대하여 서술한 바 있다. 본 논문에서는 전처리 된 영상 데이터를 이용하여 추출해 낼 수 있는 선충의 형태적 특징들을 제시한다. 선충의 크기와 관련한 특징과 자세에 관련한 특징으로 나누어, 각 특징의 추출 알고리즘을 수학적으로 표현하였다. 실험에서 제시된 형태적 특징 정보를 이용하여 직접 분류해 봄으로써 성능을 확인하였다. 분류 알고리즘은 Hierarchical Clustering을 사용하였다. 그 결과 실험에 이용된 선충의 4 종류 모두 90% 이상 옳게 분류되었다.