• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.188-193
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• 1995

A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-$\alpha$-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both $2-deoxy-D-[U-^{14}C]glucose{\;}and{\;}methyl-{\alpha}-D-[U-^{14}C]glucopyranoside$ could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.582-588
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• 1999

A Brevibacterium lactofermentum gene coding for a glucose-specific permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned, by complementing an Escherichia coli mutation affecting a ptsG gene with the B. lactofermentum genomic library, and completely sequenced. The gene was identified as a ptsG, which enables an E. coli transformant to transport non-metabolizable glucose analogue 2-deoxyglucose (2DG). The ptsG gene of B. lactofermentum consists of an open reading frame of 2,025 nucleotides encoding a polypeptide of 674 amino acid residues and a TAA stop codon. The 3' flanking region contains two stem-loop structures which may be involved in transcriptional termination. The deduced amino acid sequence of the B. lactofermentum enzyme $II^{GIe}$ specific to glucose ($EII^{GIe}$) has a high homology with the Corynebacterium glutamicum enzyme $II^{Man}$ specific to glucose and mannose ($EII^{Man}$), and the Brevibacterium ammoniagenes enzyme $II^{GIc}$ specific to glucose ($EII^{GIc}$). The 171-amino-acid C-terminal sequence of the $EII^{Glc}$ is also similar to the Escherichia coli enzyme $IIA^{GIc}$ specific to glucose ($IIA^{GIc}$). It is interesting that the arrangement of the structural domains, IIBCA, of the B. lactofermentum $EII^{GIc}$ protein is identical to that of EIIs specific to sucrose or $\beta$-glucoside. Several in vivo complementation studies indicated that the B. lactofermentum $EII^{Glc}$ protein could replace both $EII^{ Glc}$ and $EIIA^{Glc}$ in an E. coli ptsG mutant or crr mutant, respectively.

• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.292-300
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• 2003

Pseudomonas fluorescens B16 is a plant glowth-prornoting rhizobacterium, which produces an antibacterial compound that is effective against plant root pathogens, such as Agrobacrerium tumefaciens and Raistonia solanacearum. We mutagenized the strain B16 with Omegon-Km and isolated six antibacterial-activity-deficient mutants. Two cosmid clones that hybridized with the mutant clones also were isolated from a genomic library of tile parent strain. Using deletion and complementation analyses, it was found that the biosynthesis genes resided in a 4.3-kb SalI-NarI fragment. When a plasmid clone carrying the fragment was introduced into P. fluorescens strain 1855.344, which does not exhibit any antibacterial activity, the transconjugants exhibited antibacterial activity, indicating that the plasmid clone carried all the genes essential for production of the antibacterial compound. DNA sequence analysis of the fragment identified four putative open reading frames (ORFs): orf1 through orf4 The deduced amino acid sequences of ORF1, ORF2, and ORF4 were similar to cystathionine gamma lyase, pyruvate formate-lyase activating enzyme, and transcriptional regulator, respectively, yet the amino acid sequence of ORF3 showed no similarities to any known proteins. It was also demonstrated that the antibacterial activity was responsible for biological control of the bacterial wilt caused by R. solanacearum.

• 한국콘텐츠학회논문지
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• 제17권9호
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• pp.375-384
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• 2017

본 연구의 목적은 국내외 뮤지컬관련학과의 교육과정을 비교분석하고 이를 토대로 우리나라 교육과정의 문제점과 보완점이 무엇인지 제시하는데 있다. 이에 본 연구에서는 국내외의 교과과정의 차이를 비교하기기 위하여 4년제와 2~3년제 국내 대학의 뮤지컬관련학과와 국외로는 미국의 뮤지컬관련 학교 5개교, 프랑스 5개교를 선정하여 무용교육과정 수업실태를 연구하였다. 그 결과 우리나라의 뮤지컬 발전을 위해서는 교육과정의 개편이 필요하다는 결론을 도출 할 수 있었다. 첫째, 뮤지컬 현장에 맞는 체계적인 무용교육과정이 필요하다. 둘째, 다양화된 뮤지컬의 표현에 맞는 무용테크닉 수업을 강화해야 한다. 셋째, 특성화된 뮤지컬무용교과 교수법이 필요하다. 무엇보다 현장과 연계된 효율적인 교육방안이 필요하다. 뮤지컬을 공부하는 학생들은 연기, 노래, 무용을 동시에 수행해야하는 어려움에 직면해 있다. 이들 중 한 분야를 습득하기에도 어려운 것이 현실이다. 누구도 흉내 낼 수 없는 독창적인 한국스타일의 뮤지컬이라는 숙제를 해결하기 위해 대학에서의 체계적인 교육과정이 매우 중요하다.

• Journal of Microbiology and Biotechnology
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• 제23권1호
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• pp.15-21
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• 2013

We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

• Journal of Microbiology and Biotechnology
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• 제27권9호
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• pp.1692-1700
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• 2017

Ralstonia solanacearum causes bacterial wilt in a wide variety of host plant species and produces a melanin-like blackish-brown pigment in stationary phase when grown in minimal medium supplemented with tyrosine. To study melanin production regulation in R. solanacearum, five mutants exhibiting overproduction of melanin-like pigments were selected from a transposon (Tn) insertion mutant library of R. solanacearum SL341. Most of the mutants, except one (SL341T), were not complemented by the original gene or overproduced melanins. SL341T showed Tn insertion in a gene containing a conserved domain of eukaryotic transcription factor. The gene was annotated as a hypothetical protein, given its weak similarity to any known proteins. Upon complementation with its original gene, the mutant strains reverted to their wild-type phenotype. SL341T produced 3-folds more melanin at 72 h post-incubation compared with wild-type SL341 when grown in minimal medium supplemented with tyrosine. The chemical analysis of SL341T cultural filtrate revealed the accumulation of a higher amount of homogentisate, a major precursor of pyomelanin, and a lower amount of dihydroxyphenylalanine, an intermediate of eumelanin, compared with SL341. The expression study showed a relatively higher expression of hppD (encoding hydroxyphenylpyruvate dioxygenase) and lower expression of hmgA (encoding homogentisate dioxygenase) and nagL (encoding maleylacetoacetate isomerase) in SL341T than in SL341. SL341 showed a significantly higher expression of tyrosinase gene compared with SL341T at 48 h post-incubation. These results indicated that R. solanacearum produced both pyomelanin and eumelanin, and the novel hypothetical protein is involved in the negative regulation of melanin production.

• 한국조명전기설비학회:학술대회논문집
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• 한국조명전기설비학회 2009년도 춘계학술대회 논문집
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• pp.408-411
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• 2009

In this paper, conventional PI, fuzzy neural network(FNN) and adaptive teaming mechanism(ALM)-FNN for rotor field oriented controlled(RFOC) induction motor are studied comparatively. The widely used control theory based design of PI family controllers fails to perform satisfactorily under parameter variation nonlinear or load disturbance. In high performance applications, it is useful to automatically extract the complex relation that represent the drive behaviour. The use of learning through example algorithms can be a powerful tool for automatic modelling variable speed drives. They can automatically extract a functional relationship representative of the drive behavior. These methods present some advantages over the classical ones since they do not rely on the precise knowledge of mathematical models and parameters. Comparative study of PI, FNN and ALM-FNN are carried out from various aspects which is dynamic performance, steady-state accuracy, parameter robustness and complementation etc. To have a clear view of the three techniques, a RFOC system based on a three level neutral point clamped inverter-fed induction motor drive is established in this paper. Each of the three control technique: PI, FNN and ALM-FNN, are used in the outer loops for rotor speed. The merit and drawbacks of each method are summarized in the conclusion part, which may a guideline for industry application.

• 한국균학회지
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• 제38권1호
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• pp.29-33
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• 2010

KGD1 유전자는 비허용온도에서 세포벽에 결함을 보이는 Saccharomyces cerevisiae LP0353 균주의 베타-1,3-글루칸 합성 효소의 활성을 회복시키는 유전자로 분리되었다. $\alpha$-ketoglutarate dehydrogenase를 암호화하는 KGD1 유전자의 효모의 세포벽 합성과 연관된 기능을 분석하기 위하여 유전자 파괴를 시도하였다. KGD1돌연변이는 생장속도가 감소하고, 키틴 합성 효소들의 활성이 증가하였으며, 세포벽 구성 당류의 함량에 변화를 보였다. 또한 Calcofluor white과 Nikkomycin Z 등과 같은 세포벽 합성 저해물질에 대해 감수성 변화를 나타냈다. 이러한 결과들은 KGD1이 효모의 세포벽 특히 베타-1,6-글루칸과 키틴의 생합성에 영향을 주고 있음을 시사한다.

• The Plant Pathology Journal
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• 제27권1호
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• pp.89-92
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• 2011

Cyclic AMP receptor-like protein (Clp), is known to be a global transcriptional regulator for the expression of virulence factors in Xanthomonas campestris pv. campestris (Xcc). Sequence analysis showed that Xanthomonas oryzae pv. oryzae (Xoo) contains a gene that is strongly homologous to the Xcc clp. In order to determine the role of the Clp homolog in Xoo, a marker exchange mutant of $clp_{xoo4158}$ was generated. Virulence and virulence factors, such as the production of cellulase, xylanase, and extracellular polysaccharides (EPS) and swarming motility were significantly decreased in the $clp_{xoo4158}$ mutant. Moreover, the mutation caused the strain to be more sensitive to hydrogen peroxide and to over-produce siderophores. Complementation of the mutant restored the mutation-related phenotypes. Expression of $clp_{xoo4158}$, assessed by reverse-transcription realtime PCR and clp promoter activity, was significantly reduced in the rpfB, rpfF, rpfC, and rpfG mutants. These results suggest that the clp homolog, $clp_{xoo4158}$, is involved in the control of virulence and resistance against oxidative stress, and that expression of the gene is controlled by RpfC and RpfG through a diffusible signal factor (DSF) signal in Xanthomonas oryzae pv. oryzae KACC10859.

• 미생물학회지
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• 제38권3호
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• pp.173-179
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• 2002

발광 박테리아인 Photobacterium species의 lux 오페론에서 발견된 riboflavin 생합성에 관여하는 유전자들(ribI,II,III,IV)의 기능을 조사하였다. 대장균에서 이들 유전자가 포함된 재조합 플라스미드를 발현시켰을 때 상당량의riboflavin이 합성되는 것을 확인하였으며, 또한 이들 유전자들(ribI,II,III,IV)의 기능을 riboflavin에 대하여 종속 영양체인 대장균 돌연변이주(BSV 11,18)를 이용한 유전학적인 방법과 생화학적 방법으로 분석한 결과, 이들은 각각 riboflavin synthase, 3,4-dihydroxy-2-butanone 4-phosphate (DHBP) synthase, lumazine synthase, GTP cyclohydrolase II활성도를 갖는 단백질을 코드하는 것으로 밝혀졌다. 이는Photobacterium species의 riboflavin 유전자 체계가 riboflavin 생합성에 관여하는 모든 5개의 유전자들이 한 오페론에 존재하는 Bacillus subtilis와 주요 riboflavin 유전자들이 분리되어 있는 대장균과는 다른, 중간적인 형태를 갖는다는 것을 나타낸다.