• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.206-216
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• 2015

To clarify the role of surface protective antigen A (SpaA) in the pathogenesis of Erysipelothrix rhusiopathiae C43065 (serotype 2), the spaA deletion mutant of E. rhusiopathiae ${\Delta}spaA$ was constructed by homologous recombination. The virulence of the ${\Delta}spaA$ mutant decreased more than 76-fold compared with that of the wild-type strain C43065 in mice. The mutant strain was sensitive to the bactericidal action of swine serum, whereas the wild-type strain was resistant. The adhesion of wild-type strain to MEF cells was inhibited significantly by treatment with rabbit antiserum against recombinant SpaA (rSpaA) as compared with the treatment with normal rabbit serum, but the mutant strain was not affected. The mutant strain was readily taken up by mouse peritoneal macrophages in the normal rabbit serum, whereas the wild-type strain was resistant. Whereas the rabbit antiserum against rSpaA promoted the phagocytosis of wild-type strain by macrophages, the mutant strain was not affected. In addition, mice vaccinated with the formalin-killed mutant strain were provided 40% protection against challenge by the homologous virulent strain as compared with those with wild-type strain, NaOH-extracted antigen, or rSpaA, which provided more than 80% protection against the same infection. These suggested that SpaA has an important role in the pathogenesis of E. rhusiopathiae infection and could be a target for vaccination against swine erysipelas.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.184-188
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• 1987

Evidences that the mannose permease of Escherichia coli mediates the infection of N4 in early steps, were obtained as follows. First, A mutant strain of Escherichia coli which was resistant to both wild type N4 and lambda whose genome is Charon 4A containing human genomic fragments in its EcoR I site, could not use mannose efficiently. Second, N4 could not infect pel mutant strains which lack one or all of intact components of mannose permease. However, unknown alterations in N4 made it possible for the phage to infect pel mutant of E. coli. It also turned out to be clear that the receptor of N4 was different from that of lambda.

• BMB Reports
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• v.31 no.3
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• pp.287-295
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• 1998

Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of L-Ieucine, L-isoleucine, and L-valine. The sulfonylurea-resistant ALS gene from Nicotiana tabacum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS3 was used to transform Escherichia coli strain XL1-Blue, and the mutant tobacco ALS (mALS) was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). The fusion product GST-mALS was purified in a single step on a glutathione-Sepharose column. ALS activities of 0.9-2.5 ${\mu}mol/min/mg$ protein were observed in the GST-mALS, and the Km values for pyruvate, FAD, and TPP were 10.8-24.1, $(1.9-8.9){\times}10^{-3}$, and 0.14-0.38 mM, respectively. The purified GST-mALS was resistant to both the sulfonylurea and the triazolopyrimidine herbicides, and lost its sensitivity to end products, L-valine and L-leucine. For comparision, the tobacco wild-type recombinant ALS fused with GST, GST-wALS, was also characterized with respect to its pyruvate and cofactor bindings. These results suggest that the purified mutant recombinant tobacco ALS was functionally active, that the mutations resulting in herbicide resistance has affected pyruvate and cofactor bindings," and that the two classes of herbicides interact at a common site on the plant ALS.

• Microbiology and Biotechnology Letters
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• v.28 no.2
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• pp.63-70
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• 2000

To determine an increased acid tolerance of an adipic acid-resistant mutant Leuconostoc mesenteroides(ANaM100) developed for use as a Kimchi starter, proton permeability of cytoplasm, activities of H+-ATPase, Mg++ release and fatty acid composition of cytoplasmic membranes of strain ANaM100 were studied and compared with those of its wild type (LMw). The value of protons permeability of LMw after an acid shock at pH 5.0 was 5.4 min., while the value of ANaM100 cells was 8.4 min. at the same pH. The pH of maximal specific activ-ities of ATPase originated from the LMw and ANaM100 were 0.87 unit/mg protein at pH 6.0 and 0.92 unit/mg pro-tein at pH 5.5, respectively. The release of magnesium ion from ANaM100 was observed about 12.8% at pH 4 after 2 hours, while the wild strains of LMw released Mg++ about 27.6% under the same conditions. The content of C19:0,cyclo and C18:1 in a membrane fatty acid of ANaM100 was higher and lower, respectively than that of LMw. These results indicated that acid tolerance of adipic acid-resistant strain, ANaM100 was significantly improved in comparison with that of its wild type, LMw. In addition, the strain ANaM100 was adipic resistance based on the result of growth of the strain in comparison with that of strain LMw in a broth containing adipic acid.

• Journal of Microbiology and Biotechnology
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• v.28 no.8
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• pp.1384-1390
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• 2018

Glyphosate inhibits the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in the shikimate pathway. A mutant of EPSPS from Pantoea sp. was identified using site-directed mutagenesis. The mutant showed significantly improved glyphosate resistance. The mutant had mutations in three amino acids: Gly97 to Ala, Thr 98 to Ile, and Pro 102 to Ser. These mutation sites in Escherichia coli have been studied as significant active sites of glyphosate resistance. However, in our research, they were found to jointly contribute to the improvement of glyphosate tolerance. In addition, the level of glyphosate tolerance in transgenic Arabidopsis confirmed the potentiality of the mutant in breeding glyphosate-resistant plants.

• The Journal of Natural Sciences
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• v.8 no.2
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• pp.43-48
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• 1996

Mutangen sensitive hyper-rec type musN mutants were assigned into the UvsC group which contains genes involved in recombination and mutation. however, phenotypic properties of musN mutants were very different from those found in uvsC mutant strains which are rec- and lack UV-induced mutation. musN was not a mutator like uvsC. In addition, selenate resistant mutations in musN were induced sililar to those in wild types by UV irradiation. Wild type levels of UV-sensitivity in dividing cells of musN also differ from the uvsC phenotypes. These indicate that the UvsC group has branched pathways.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.581-584
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• 2019

In this study, the carbon monoxide (CO)-fermenting acetogen, Clostridium sp. AWRP was subjected to chemical mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) to generate a CO-resistant mutant. Among the 26 colonies obtained, the highest alcohol production was observed in one isolate, named C1. Compared to the wild-type strain, the C1 strain exhibited 1.5- and 3.4-fold higher CO consumption rate and alcohol selectivity, respectively. The total CO consumption of strain C1 could be further enhanced by increasing the content of metal ions, such as nickel and iron. The highest ethanol titer (5.7 g/l) was achieved by 5-fold increase in the iron concentration.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.121-128
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• 1997

A lactic acid bacterium was isolated from kimchi. The isolated strain was identified as the genus Lactobacillus through its morphological, cultural, and physiological characteristics and named as Lactobacillus sp. JC-7. The optimum conditions for the electrofusion between streptomycin (2.5 mg/ml) resistant mutant of Lactobacillus acidophilus 88 and kanamycin (600 $\mu$g/ml) resistant mutant of Lactobacillus sp. JC-7 were evaluated. The highest number of fusants were obtained at a capacitance value of 120 msec (1670 $\mu$F), a field strength of 100 V/cm, and a pulse controller setting of 72$\Omega$. The optimum pH of electroporation buffer was 7.5 and the concentration of divalent cation was 1 mM Mg$^{2+}. Electrofusants were efficiently obtained by addition 20% polyethylene glycol to elec- troporation buffer. The yield of fusion was better than that of using polyethylene glycol mediated chemical induction.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.68.1-68
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• 2003

When plants are infected by plant pathogens, typical disease symptom termed lesion, appears in compatible interaction. Whereas, in incompatible interactions, only small speck of lesions are visible on the leaf surfaces. Hypersensitive response (HR) of plant which is the result of infection by incompatible pathogens, is a well known defense response inducing rapid cell death resulting in complete resistance. However, some rice mutants show spontaneous disease symptoms during the growth stages without interaction with pathogens. We investigated the spontaneous cell death mutant called Blast Lesion Mimic(BLM) generated by EMS mutation, on the relationship with the hypersensitive response as well as resistant characteristics. Accumulation of phenolic compounds were detected around the lesions as lesions develop on leaf surface. Activation of PR gene was detected before the lesion appeared, and that result indicates the defense-related response are started earlier than lesion formation. The BLM mutant showed resistant response to inoculation of Magnaporthe grisea KJ201 with which the wild type Hwacheong is totally susceptible. Informations on the formation of spontaneous lesions and detail analysis of lesion mimic mutants and related genes are very limited to date. It is really important to understand the phenomenon of the defense-related lesion formation for developing resistant cultivar for rice blast pathogens

• Korean Journal of Veterinary Research
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• v.41 no.3
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• pp.343-350
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• 2001

The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.