• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.247-256
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• 1999

The present study was performed to improve the reproductive disturbance as well as the elimination of microbiological contamination for animals bred under conventional conditions followed by in vitro fertilization and embryo transfer techniques including embryo and sperm freezing, using a mouse strain(M. m. molossinus-tt＠Kist) showing the abnormal behavior disorder derived from Korean wild mice (Mus musculus molossinus). Moreover, hematological and serum biochemical analyses were also carried out to obtain the basic data of this mouse strain The results are summarized as follows: 1. In comparison with hematological data, the numbers of RBC and platelet of this mouse strain were appeared as the higher value those that of the same aged inbred strains such as BALB/c, DBA/2, C57BL/6 and C3H /Hen. However, no differences were found in values of WBC, Hb and Ht. Moreover, total cholesterol of this strain showed a low value but triglyceride, total protein and albumin values were similar as in inbred strains. 2. The average numbers of superovulated oocytes treated with 2.5/2.5 IU and 5.0/5.0 IU of PMSG/hCG were 11.6 and 12.7, respectively. The fertilization rates of 2.5/2.5 IU PMSG /hCG treatment(87.9%) was higher than 5.0/5.0 IU treatment(52.0%) (p＜0.05) and the developmental rate of 2 cell stage embryos were 외 so appeared as higher value 99.0% and 90.6%, respectively. 3. The rates of in vitro fertilization treated with frozen sperm(24.8%) was significantly lower than of that fresh sperm(87.9%), (p＜0.05). 4. The five, six and ten heads of offspring were obtained from frozen-thawed 2 cell embryos by in vitro fertilized, 2 cell embryos from in vitro fertilized by frozen-thawed spermatozoa. and 2 cell embryos by in vitro fertilization, respectively. These offspring developed the expected disease about 2 weeks after birth, which was confirmed that the disease character of this mutant mouse strain was reliably reproduced. 5. MHV(Mouse hepatitis virus) and Staphylococcus aureus were successfully eliminated from conventional animals by in vitro fertilization-embryo transfer and the use of SPF recipient animals.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.454-458
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• 2006

HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

• Journal of Life Science
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• v.18 no.11
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• pp.1606-1611
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• 2008

Outer membrane proteins (OMPs) expressed in the Gram negative bacteria such as Salmonella play multiple functions including material transports, adhesive factors and reception of external signals. This study has been focused on an OmpW protein known as a protein required to form a hydrophobic porin in outer membrane. We have constructed a S. typhimurium CK10 mutant deleting an ompW gene on chromosome. The CK10 strain was more tolerant to SDS than the wild-type strain did. As increase of salt concentration in the culture media, significantly decreased amount of OmpW protein in cells were detected. The maximum OmpW protein was expressed in the absence of salt supplement. However, the growth of CK10 strain was indistinguishable compared to that of the wild-type strain at the variable osmotic conditions. The biological role of differential OmpW expression in response to osmotic conditions remains to be investigated.

• Food Science and Preservation
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• v.22 no.6
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• pp.908-914
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• 2015

The objectives of this study were to isolate and characterize low temperature adaptation yeast and to obtain suitable yeasts strains for manufacturing Yakju. In this study, we isolated 482 wild yeasts from fermented foods. Out of these, 5 yeast strains were selected based on increased growth at low temperature ($15^{\circ}C$) and high ${\beta}$-glucosidase activity. To screen the aromatic level of isolates, media containing cerulenin and 5,5,5-trifluor-DL-leucine (TFL) were used. Y297 strain demonstrated tolerance against TFL and produced more than 13% alcohol. Y297 strain was identified a Saccharomyces cerevisiae based on the 26S rDNA gene sequences. Maximum cell growth was observed after 19 hr and 38 hr of incubation at $25^{\circ}C$ and $15^{\circ}C$, respectively. The exponential phase was followed by a lengthy stationary phase, at $15^{\circ}C$, when the cells remained high viable. Y297 strain demonstrated tolerance against alcohol (10%), glucose (60%) and salt(NaCl, 8%). ${\beta}$-glucosidase and esterase activity in Y297 were higher than those of controls at $15^{\circ}C$. Overall, these results indicated that using wild yeast strain, isolated from fermented food, affects the chemical characteristics of the brewed Yakju.

• Clean Technology
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• v.5 no.2
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• pp.79-86
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• 1999

Mutant strain EID was developed by treating Gordona sp. CYKS1 with ethylmethanesulfone, and the desulfurization characteristics of dibenzothiophene(DBT) by mutant EID was investigated. Strain EID desulfurized DBT to 2-hydroxybiphenyl (2-HBP) by 4S pathway. Desulfurization rate of the strain EID was $4.0{\mu}mol{\cdot}L^{-1}{\cdot}h^{-1}$, while that of the wild type CYKS1 was $2.6{\mu}mol{\cdot}L^{-1}{\cdot}h^{-1}$. The effect of glucose concentration supplied as the carbon source on the DBT desulfurization showed that DBT desulfurization rate was enhanced as the glucose concentration increased. Maximum DBT desulfurization rate was $11.1{\mu}mol{\cdot}L^{-1}{\cdot}h^{-1}$ at 2.0 mM DBT concentration. As end-products such as 2-HBP and sulfate concentrations increase, DBT desulfurization activity of the strain EID decreased. When 0.2 mM of 2-HBP was added in the medium, no growth and desulfurization activity was observed. When 0.5 g/L $Na_2SO_4$ was simultaneously supplied with DBT, DBT desulfurization rate was$1.4{\mu}mol{\cdot}L^{-1}{\cdot}h^{-1}$.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.266-271
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• 2018

Effect of plasmid curing of Acinetobacter sp. B-W on the production of siderophore from glutamic acid as both carbon and nitrogen sole sources was investigated. Plasmid cured mutant of strain B-W lost the ability to produce siderophore from glutamic acid at $28^{\circ}C$. Transformant E. coli $DH5{\alpha}$ harboring 20 kb plasmid, that was isolated from wild type of strain B-W produced siderophore from glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$, but, not at $36^{\circ}C$. Production of siderophore from glutamic acid by transformant E. coli $DH5{\alpha}$ was completely inhibited by $10{\mu}M\;FeCl_3$. In previous report, catechol nature of siderophore produced from glutamic acid by strain B-W was detected by Arnow test. The siderophore produced from glutamic acid by transformant E. coli $DH5{\alpha}$ was also catechol type. Rf value of siderophore produced from transformant E. coli $DH5{\alpha}$ grown in medium glutamic acid as both carbon and nitrogen sole sources at $28^{\circ}C$ was 0.32 in butanol-acetic acid-water (12:3:5) as developing solvent. Rf value of the siderophore was the same with that of wild type of strain B-W. Thus a single plasmid of 20 kb seemed to be involved in the production of siderophore from glutamic acid.

• Journal of Microbiology and Biotechnology
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• v.29 no.11
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• pp.1806-1816
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• 2019

Candida albicans is an opportunistic fungus possessing multiple virulence factors controlling pathogenicity. Cell wall proteins are the most important among these factors, being the first elements contacting the host. Ddr48 is a cell wall protein consisting of 212 amino acids. A DDR48 haploinsufficient mutant strain was previously found necessary for proper oxidative stress response and drug resistance. In this study, we aimed to further elucidate the role of Ddr48 by performing additional phenotypic characterization assays. A combinatory proteomic and bioinformatics approach was also undertaken to determine differentially expressed cell wall proteins. Results showed that the mutant strain exhibited a 10% decrease in adhesion mirrored by a 20% decrease in biofilm formation, and slight sensitivity to menadione, diamide, and SDS. Both strains showed similar hyphae formation, virulence, temperature tolerance, and calcofluor white and Congo red sensitivities. Furthermore, a total of 8 and 10 proteins were identified exclusively in the wild-type strain grown under filamentous and non-filamentous conditions respectively. Results included proteins responsible for superoxide stress resistance (Sod4 and Sod6), adhesion (Als3, Hyr4, Pmt1, and Utr2), biofilm formation (Hsp90, Ece1, Rim9, Ipp1, and Pra1) and cell wall integrity (Utr2 and Pga4). The lack of detection of these proteins in the mutant strain correlates with the observed phenotypes.

• The Plant Pathology Journal
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• v.33 no.3
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• pp.276-287
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• 2017

RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.

• Food Science of Animal Resources
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• v.41 no.6
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• pp.967-982
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• 2021

Probiotics are living microorganisms that, when administered in adequate amounts, provide a health benefit to the host and are considered safe. Most probiotic strains that are beneficial to human health are included in the "Lactic acid bacteria" (LAB) group. The positive effects of probiotic bacteria on the host's health are species-specific and even strain-specific. Therefore, evaluating the probiotic potential of both wild and novel strains is essential. In this study, the probiotic characteristics of Lactobacillus brevis KT38-3 were determined. The strain identification was achieved by 16S rRNA sequencing. API-ZYM test kits were used to determine the enzymatic capacity of the strain. L. brevis KT38-3 was able to survive in conditions with a broad pH range (pH 2-7), range of bile salts (0.3%-1%) and conditions that simulated gastric juice and intestinal juice. The percentage of autoaggregation (59.4%), coaggregation with E. coli O157:H7 (37.4%) and hydrophobicity were determined to be 51.1%, 47.4%, and 52.7%, respectively. L. brevis KT38-3 produced β-galactosidase enzymes and was able ferment lactose. In addition, this strain was capable of producing antimicrobial peptides against the bacteria tested, including methicillin and/or vancomycin-resistant bacteria. The cell-free supernatants of the strain had high antioxidant activities (DPPH: 54.9% and ABTS: 48.7%). Therefore, considering these many essential in vitro probiotic properties, L. brevis KT38-3 has the potential to be used as a probiotic supplement. Supporting these findings with in vivo experiments to evaluate the potential health benefits will be the subject of our future work.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1211-1216
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• 2004

A MNNG (N-methyl-N'-nitro-N-nitrosoguanidine) induced chromium resistant strain ($Cr^{r}18$) of unicellular cyanobacterium Anacystis nidulans has been isolated and characterized. The resistant strain could grow (although restricted to $50\%$ of control) in chromium concentration (180${\mu}M$) lethal to the wild-type. Sublethal ($160{\mu}M$) concentration of $Cr^{6+}$ significantly reduced (13-$37.5$) all the photosynthetic pigments of A. nidulans with maximum reduction in phycoerythrin followed by ChI $\alpha$. Pigments of A. nidulans were drastically decreased in lethal concentration of Cr^{6+} with maximum reduction in phycoerythrin ($75\%$) and allophycocyanin ($67.5\%$). Resistant strain $Cr^{r}18$ resisted toxic effects of sublethal and lethal concentrations of $Cr^{6+}$ on photosynthetic pigments as revealed by less decrease in pigments as compared to A. nidulans. Effect of $Cr^{6+}$ stress was also studied on nitrogen assimilation and phosphate uptake. Sublethal concentration of $Cr^{6+}$ drastically reduced ($71.5\%$) nitrate uptake by A. nidulans while a decrease of $29\%$ was observed in strain $Cr^{r}18$. Short (2 day) exposure of A. nidulans and its resistant strain $Cr^{r}18\;to\;Cr^{6+}$ did not affect nitrate reductase and glutamine synthetase (transferase), whereas longer (10 day) exposure to $Cr^{6+}$ lowered activities of both enzymes in A. nidulans but not significantly in the strain $Cr^{r}18$. Ammonium uptake by both strains was not affected by $Cr^{6+}$. Thus, $Cr^{6+}$ affected photosynthetic pigments, nitrogen assimilation, and phosphate uptake of A. nidulans, while strain $Cr^{r}18$ was able to resist toxic effects of the metal. Advantages of using strain $Cr^{r}18$ for bioremediation purposes have been evaluated by studying $Cr^{6+}$ removal from the solution. Resistant strain $Cr^{r}18$ was able to remove $33\%$ more $Cr^{6+}$ than A. nidulans and thus it can prove to be a good candidate for bioremediation of $Cr^{6+}$ from polluted waters.