• Microbiology and Biotechnology Letters
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• 제18권5호
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• pp.535-540
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• 1990

Bacitlus sphaericus ts-Dl290 was characterized comparatively with the wild type strain 1593 by themeasurements of the biosynthesis of total DNA, RNA and protein on the temperature-shift culturesat permissive temperature of $30^{\circ}C$ and at nonpermissive temperature of $42^{\circ}C$. The growth patterns of the wild type strain and ts-Dl290 were similar at $30^{\circ}C$, but at 4Z C the mutant almost did not grow (temperature-sensitivity). When the growth temperatures of both stains were shifted-up from $30^{\circ}C$ to $42^{\circ}C$ after a 4 hour culture, their growths were normal, but when shifted-down from $42^{\circ}C$ to $30^{\circ}C$ after a 4 h culture, the mutant did not grow. When shifted up from $30^{\circ}C$ to $42^{\circ}C$ after a 4 hculture, the DNA syntheses of the two strains were at a normal rate for 1 h, but after 1 h the biosynthesesdecreased. The rate of DNA synthesis of the wild type strain at the nonpermissive temperature was about 93%, and that of the mutant was about 50% of the ratio of the wild type strain, and the RNA synthesis of the wild type strain was maintained for 3 h, and that of the mutant for 2 h. Thereafter the RNA synthesis decreased, and the synthesis of proteins in the both strains were similarlykept high for 8 h. The reversibility of the DNA synthesis of the mutant at $42^{\circ}C$ was lessened whenthe culture times were increased.re times were increased.

• Journal of Life Science
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• 제18권2호
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• pp.148-153
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• 2008

Phytochrome double mutant (PhyAB) showed the delayed root gravitropic response compared to the wild type (WT) in Arabidopsis. After 8 hr of gravistimulation, the gravitropic response of mutant showed 48% of the WT. The delayed response started at 1.5 hr after gravistimulation. And we measured the ethylene production in the root segments of WT and mutant for 12 hr. Ethylene production of mutant decreased about 40% of the WT at 12 hr. This result suggested that the phytochrome might be linked with ethylene production in some way. Generally, ethylene inhibits the growth of plant organs including roots. We measured the root growth rate in the presence of ACC (1-aminocyclopropane-1-carboxylic acid), a precursor of ethylene. And WT showed the inhibition of root growth with ACC, but mutant did not show the inhibition as WT did. To confirm the relationship between the ethylene and gravitropic response, we measured the gravitropic response with ACC. In the presence of $10^{-6}$ M ACC, WT showed the 37.4% inhibition compared to the control (no ACC), whereas mutant showed the only 6.6% inhibition of control (no ACC). This research suggested the relationship between phytochrome and gravitropic response through an ethylene production.

• Journal of Life Science
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• 제25권9호
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• pp.1051-1058
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• 2015

Citrus mutant fruits were induced by irradiation of citrus budsticks with 120 Gy of cobalt (⁶⁰CO) gamma irradiation. The citrus mutant inhibited the growth and induced apoptosis in various human cancer cells, including A549, HepG2, HCT116, MCF-7, and Hela. The results of a trypan blue exclusion assay showed that citrus mutant fruits exhibited excellent antiproliferation activity in various human cancer cells and low cytotoxicity in normal 16HBE140- and CHANG cells. In addition, the cell death induced by the citrus mutant fruits was associated with an increased population of cells in sub-G1 phase, and it caused DNA fragmentation in human lung adenocarcinoma A549 and hepatocellular carcinoma HepG2 cells. It also up-regulated the amount of cellular nitric oxide (NO^•) produced as a result of nitric oxide synthase (NOS) activation and suppressed the inhibitor of apoptosis protein (IAP) family in A549 and HepG2 cells. These findings indicate that the citrus mutant fruits activates the NO^•-mediated apoptotic pathway in A549 and HepG2 cells. It may merit further investigation as a potential chemotherapeutic and chemopreventive agent for the treatment of various types of cancer cells. The results provide important major new insights into the mechanisms of the anticancer activity of citrus mutant fruits.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1271-1279
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• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• Journal of Life Science
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• 제27권3호
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• pp.339-345
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• 2017

Carotenoids are natural lipid-soluble pigments, which are produced primarily by bacteria, algae, and plants. Many studies have focused on the identification, production, and utilization of natural sources of astaxanthin from algae, yeast, and crustacean byproducts as an alternative to the synthetic pigment, which is mostly used today. The aim of the present study was to identify a mutant of Paracoccus haeundaensis by exposure to UV and ethyl methanesulfonate (EMS). The mutant was then exposed to nutrient stress conditions to isolate an astaxanthin-hyperproducing strain, followed by characterization of the mutant. The survival rate decreased in accordance with an increase in the UV exposure time and an increase in the EMS concentration. A mutant of the original P. haeundaensis strain was identified that showed hyperproduction of astaxanthin following exposure to UV irradiation (20 min) and EMS treatment (0.4 M concentration). The optimal culture conditions for the PUE mutant were $25^{\circ}C$, pH 7-8, and 3% NaCl. The effects of various carbon and nitrogen sources on the growth and astaxanthin production of PUE were examined. The addition of 1% raffinose and 3% potassium nitrate influenced cell growth and astaxanthin production. The selected mutant exhibited an increase of 1.58 folds in astaxanthin content compared to initial wild type strain. A genetically stable mutant strain obtained using mutagen (UV irradiation and EMS treatment) may be a suitable candidate for further industrial scale production of astaxanthin.

• Korean Journal of Microbiology
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• 제37권2호
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• pp.170-175
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• 2001

In order to produce xylitol with high yield, experiments were carried out to develope xylitol dehydrogenase(XDH) defective mutant from P. stipitis and to investigate the xylitol fermentation characteristics of mutant strain. After treatment of P. stipitis with EMS, mutant PXM-4 was selected based on te XDH activity and xylitol production capability. Among the tested cosubstrates, galactose was selected as an adequate cosub-strate on xylitol production of mutant PXM-4. But with the increase in the concentration of galactose in the medium, xylitol production was decreased because the transport of xylose into cell was inhibited by galactose. The optimal concentration of galactose for the production of xylitol using 20 g/ι xylose was 20 g/ι Under this condition, maximum concentration of xylitol and yield were 14.4 g/ι and 97%, respectively. In order to prevent the inhibitory effect of xylose transport by galactose, galactose was fed with low concentration and the concentration of xylitol produced was increased up to 25 g/ι.

• Korean Journal of Agricultural Science
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• 제13권2호
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• pp.279-288
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• 1986

One hundred and fifty one mutant strains were obtained from the parent strain Aspergillus oryzae MF by ultra-violet ray irradiation. Among those mutants a strain, Asp. oryzae UM-36 which hyperprodued protease, was selected and its morphological characteristics and the production of enzymes protease, ${\alpha}$-amylase, and glucoamylase on wheat bran koji and on soy-sauce koji were studied. The results obtained were as follows 1. The selected mutant showed slower growth and weak sporulation on malt agar and on Czapek agar than the parent strain. 2. The conidiophores of the mutant were generally shorter than those of the parent when grown on malt agar. 3. Sectoring in colonies was not found when grown on malt agar and on Czapek agar. 4. The level of protease production by the mutant was increased approximately 1,4-fold higher on wheat bran koji and 2-fold higher on soysauce koji than by the parent. 5. The production of ${\alpha}$-amylase and glucoamylase by the mutant were also increased as compared with the parent on wheat bran koji and on soy sauce koji. 6. In the case of parent strain and mutant strain, the highest activity of protease appeared after three days in wheat bran medium at $30^{\circ}C$ incubation, but the highest activities of ${\alpha}$-amylase and glucoamylase appeared after two days.

• Korean Journal of Veterinary Research
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• 제41권2호
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• Journal of Life Science
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• 제23권3호
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• pp.347-354
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• 2013

Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.

• Korean Journal of Microbiology
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• 제54권4호
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• pp.343-353
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• 2018

Our previous research demonstrated the essential role of the xenB gene in stress response to RDX by using Pseudomonas sp. HK-6 xenB knockout. We have extended this work to examine the cellular responses and altered proteomic profiles of the HK-6 xenA knockout mutant under RDX stress. The xenA mutant degraded RDX about 2-fold more slowly and its growth and survival rates were several-fold lower than the wild-type HK-6 strain. SEM revealed more severe morphological damages on the surface of the xenA mutant cells under RDX stress. The wild-type cells expressed proportionally-increased two stress shock proteins, DnaK and GroEL from the initial incubation time point or the relatively low RDX concentrations, but slightly less expressed at prolonged incubation period or higher RDX. However the xenA mutant did not produced DnaK and GroEL as RDX concentrations were gradually increased. The wild-type cells well maintained transcription levels of dnaA and groEL under increased RDX stress while those in the xenA mutant were decreased and eventually disappeared. The altered proteome profiles of xenA mutant cells under RDX stress also observed so that the 27 down-regulated plus the 3 up-regulated expression proteins were detected in 2-DE PAGE. These all results indicated that the intact xenA gene is necessary for maintaining cell integrity under the xenobiotic stress as well as performing an efficient RDX degradation process.