• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.233-240
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• 2016

FK506, a widely used immunosuppressant, is a 23-membered polyketide macrolide that is produced by several Streptomyces species. FK506 high-yielding strain Streptomyces sp. RM7011 was developed from the discovered Streptomyces sp. KCCM 11116P by random mutagenesis in our previous study. The results of transcript expression analysis showed that the transcription levels of tcsA, B, C, and D were increased in Streptomyces sp. RM7011 by 2.1-, 3.1-, 3.3-, and 4.1-fold, respectively, compared with Streptomyces sp. KCCM 11116P. The overexpression of tcsABCD g enes in Streptomyces sp. RM7011 gave rise to approximately 2.5-fold (238.1 μg/ml) increase in the level of FK506 production compared with that of Streptomyces sp. RM7011. When vinyl pentanoate was added into the culture broth of Streptomyces sp. RM7011, the level of FK506 production was approximately 2.2-fold (207.7 μg/ml) higher than that of the unsupplemented fermentation. Furthermore, supplementing the culture broth of Streptomyces sp. RM7011 expressing tcsABCD genes with vinyl pentanoate resulted in an additional 1.7-fold improvement in the FK506 titer (498.1 μg/ml) compared with that observed under non-supplemented condition. Overall, the level of FK506 production was increased approximately 5.2-fold by engineering the supply of allylmalonyl-CoA in the high-yielding strain Streptomyces sp. RM7011, using a combination of overexpressing tcsABCD genes and adding vinyl pentanoate, as compared with Streptomyces sp. RM7011 (95.3 μg/ml). Moreover, among the three precursors analyzed, pentanoate was the most effective precursor, supporting the highest titer of FK506 in the FK506 high-yielding strain Streptomyces sp. RM7011.

• BMB Reports
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• v.56 no.11
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• pp.600-605
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• 2023

Intrahepatic cholangiocarcinoma (ICC) is a bile duct cancer and a rare malignant tumor with a poor prognosis owing to the lack of an early diagnosis and resistance to conventional chemotherapy. A combination of gemcitabine and cisplatin is the typically attempted first-line treatment approach. However, the underlying mechanism of resistance to chemotherapy is poorly understood. We addressed this by studying dynamics in the human ICC SCK cell line. Here, we report that the regulation of glucose and glutamine metabolism was a key factor in overcoming cisplatin resistance in SCK cells. RNA sequencing analysis revealed a high enrichment cell cycle-related gene set score in cisplatin-resistant SCK (SCK-R) cells compared to parental SCK (SCK WT) cells. Cell cycle progression correlates with increased nutrient requirement and cancer proliferation or metastasis. Commonly, cancer cells are dependent upon glucose and glutamine availability for survival and proliferation. Indeed, we observed the increased expression of GLUT (glucose transporter), ASCT2 (glutamine transporter), and cancer progression markers in SCK-R cells. Thus, we inhibited enhanced metabolic reprogramming in SCK-R cells through nutrient starvation. SCK-R cells were sensitized to cisplatin, especially under glucose starvation. Glutaminase-1 (GLS1), which is a mitochondrial enzyme involved in tumorigenesis and progression in cancer cells, was upregulated in SCK-R cells. Targeting GLS1 with the GLS1 inhibitor CB-839 (telaglenastat) effectively reduced the expression of cancer progression markers. Taken together, our study results suggest that a combination of GLUT inhibition, which mimics glucose starvation, and GLS1 inhibition could be a therapeutic strategy to increase the chemosensitivity of ICC.

• The Journal of Korean Society of Virology
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• v.26 no.2
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• pp.163-171
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• 1996

The recombinant pseudorabies virus major capsid protein (rMCP) was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. Following evaluation of the immunochemical properties of the rMCP, the immunogenicity of the recombinant subunit protiens were investigated in guinea pig and swine to obtain the preliminary guide line for the subunit vaccine using rMCP and gP50. It was proved that ultrasonication and 30% ammonium sulfate was most efficient to concentrate and purify the protein. The rMCP was safe in mice, guinea pigs and piglets. In guinea pigs, rMCP mixed with various adjuvants induced substantial degree of serum neutralizing antibody titers, but revealed incomplete protectivity against challenge. In swine, the combination of rMCP and gP50 showed the higher serum neutralizing antibody titers and cellular immune responses than rMCP alone. However, the protectivity was lower in comparison with the commercial gI-deleted inactivated vaccine. We expect these results to contribute to characterization of MCP gene of Korean isolate of PRV and to ultilize as preliminary information for prodution and evaluation of PRV recombinant subunit vaccines.

• ALGAE
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• v.19 no.3
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• pp.161-174
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• 2004

Phonological, morphological and molecular characteristics of Spatoglossum pacificum Yendo are examined. S. pacificum has an annual life cycle composed of saprophytes with apparent absence of male and female gametophytes in Korea. The seasonal growth of this species explains that the annual growth is closely related to the monthly variation of water temperature. S. pacificum has protruding reproductive structures above the outmost cortical layer. Although this observation is restricted to several species, reproductive structures on the thallus can make S. pacificurn distinguishable from S. crassum and S. lacturn. The morphogenesis of a midrib at the base of S. pacificum in this study is the same as those of Dictyopteris but different from those of S. crassum and S. lacturn, suggesting that S. pacificum is closely related to Dictyopteris. In the comparison of plastid gene sequences among species of Spatoglossum and Dictyopteris, S. pacificum is more similar to D. divaricata and D. undulate than those of S. crassum in rbcL, rbcS, psbA and psaA. This result is congruent with the anatomical characteristic of a midrib at the base of the thallus and the protrusion of reproductive organs on the thallus. The phylogenetic relationship based on these plastid genes also shows that S. pacifism is included in Dictyotpteris Glade and separated from S. crassum. We propose the new combination of Dictyopteris pacifica (Yendo) I.K. Hwang, H.S. Kim et W.J. Lee, comb. nov. based on the differences of anatomical characteristics of the midrib, the existence of reproductive organs on thallus and the molecular analyses.

• Journal of Life Science
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• v.23 no.6
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• pp.728-735
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• 2013

Plantago asiatica (Plantaginaceae) is a wind-pollinated plant that grows mainly on fields in East Asia. Starch gel electrophoresis was used to investigate the allozyme diversity and population structure of 18 populations of this species. Although the plantain populations were isolated and patchily distributed, they maintained a high level of genetic diversity; the average percentage of polymorphic loci was 57.1%, the mean number of alleles per locus was 2.07, and the average heterozygosity for 18 populations was 0.201. The combination of a predominant wind-pollinated, mix-mating reproduction, large population sizes, high gene flow between subpopulations, and a propensity for high fecundity may explain the high level of genetic diversity within populations. A direct gradient in overall genetic diversity is associated with latitude. Genetic diversity of P. asiatica is markedly decreased from $35^{\circ}3^{\prime}$ to high latitude and decreased from $35^{\circ}3^{\prime}N$ to low latitude, whereas there does not show a longitudinal gradient in genetic diversity.

• Food Science of Animal Resources
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• v.38 no.4
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• pp.829-834
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• 2018

The objective of this study was to determine the minimum enrichment time for different types of food matrix (pork, beef, and fresh-cut lettuce) in an effort to improve Escherichia coli detection efficiency. Fresh pork (20 g), beef (20 g), and fresh-cut lettuce (20 g) were inoculated at 1, 2, and 3 Log CFU/g of Escherichia coli. Samples were enriched in filter bags for 3 or 5 h at $44.5^{\circ}C$, depending on sample type. E. coli cell counts in the samples were enriched in E. coli (EC) broth at 3 or 5 h. One milliliter of the enriched culture medium was used for DNA extraction, and PCR assays were performed using primers specific for uidA gene. To detect E. coli (uidA) in the samples, a 3-4 Log CFU/mL cell concentration was required. However, E. coli was detected at 1 Log CFU/g in fresh pork, beef, and fresh-cut lettuce after 5, 5, and 3-h enrichment, respectively. In conclusion, 5-h enrichment for fresh meats and 3-h enrichment for fresh-cut lettuce in EC broth at $44.5^{\circ}C$, and PCR analysis using uidA gene-specific primers were appropriate to detect E. coli rapidly in food samples.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.388-393
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• 2017

We report an Agrobacterium-mediated transformation procedure optimized for 'Kyoho' that is a major table grapevine cultivar in Korea, and its transgenic plants with antifreeze protein gene of carrot (DcAFP). The full length of DcAFP coding region in accordance with the previous report was isolated from young leaves of carrot and recombined into a plant transformation vector. Ethylene inhibitors such as silver nitrate and aminoethoxyvinylglycin (AVG) supplemented in a co-cultivation medium distinctly increased frequency of shoot regeneration when explants were sub-cultured in a selection medium: particularly ten-fold higher in treatment with 0.1 mg/L AVG than one without ethylene inhibitor. Among various antibiotics and their concentrations, the combination of 150 mg/L cefotaxime plus 150 mg/L $Clavamox^{TM}$ was selected for elimination of Agrobacterium cells in addition to minimization of adverse effect on shoot regeneration, while 50 mg/L kanamycin monosulfate effectively suppressed regeneration of non-transgenic shoots. Applying the elucidated culture condition, we finally obtained a total of 5 transgenic 'Kyoho' plantlets with DcAFP, of which integration with the grapevine genome and transcription was confirmed by nucleic acid analyses.

• The Korean Journal of Malacology
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• v.26 no.4
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• pp.275-283
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• 2010

Many freshwater snail taxa are difficult to identify using morphological traits due to phenotypic plasticity. However, using of molecular DNA marker in combination with morphological traits can provide a reliable means for discriminating among freshwater snail taxa including cryptic species. To discriminate among Korean freshwater snail taxa and resolve their systematic relationships, wesequenced a fragment of mtDNA cytochrome oxidase I (COI) gene from 82 specimens collected from ten different sites distributed along the Korean peninsula. We identified more than seven freshwater snail taxa including cryptic species in Korea. Whereas traditional shell morphology of freshwater snails offers only weak discriminatory power for recognizing 'good' taxa, DNA sequence data provided positive and reliable identification. In addition, a major Semisulcospira clade was clearly separated from the remaining lineages observed including cryptic species. However, a phylogenetic tree inferred from the COI gene data did not fully resolve systematic relationships among pleurocerid taxa in Korea. Establishing more robust shell characteristics for identifying taxa unambiguously and hence improving traditional key shell morphology characters for freshwater snail species is an urgent requirement and will require more rigorous examination of all nominal taxa. While molecular data generated here will be useful for species identification and for describing the systematic relationships among Korean freshwater snails, further analysis will be required.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.181-186
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• 1986

A defective lambda transducing phase carrying acetyl CoA carboxylase gene (fabE) from Escherichia coli chromosome (72 min on the current linkage map) has been isolated. A restriction map of the chromosomal region from defective transducing phage was established by digestion with combination of the restriction enzymes. No cleavage site for the enzyme EcoRI was found in this region. Restriction fragments were cloned from defective transducing phage into high copy number plasmid vector pACYC184 to generate hybrid plasmids which were capable of complementation of fabE temperature sensitive mutation. We show here that the fabE gene is located on a 3.4 megadalton Bam HI-SalI fragment with a HindIII site, which lies within the 7.4 megadalton BglIIfragment, by complementation analysis.

• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1433-1442
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• 2018

To identify and quantify the effects of a combination of dietary $1{\times}10^8CFU/g$ Lactococcus lactis subsp. lactis I2 ($LI_2$) and 0.1% ${\beta}$-glucooligosaccharides (BGO) on the growth and immunity of olive flounder (Paralichthys olivaceus), a feeding experiment was conducted. Flounder ($14{\pm}0.5g$) were divided into two groups and fed control and synbiotic feeds for 8 weeks. Investigations were carried out on growth and feed utilization, innate immunity, serum biochemical parameters, intestinal lactic acid bacterial (LAB) viability, microvillus length, and changes in the expression levels of genes encoding pro-inflammatory cytokines (tumor necrosis factor $[TNF]-{\alpha}$, interleukin $[IL]-1{\beta}$, and IL-6). Results demonstrated the synbiotic diet had significantly better (p < 0.05) responses in terms of weight gain and specific growth rate, three innate immune parameters (respiratory burst, serum lysozyme, and superoxide dismutase), intestinal LAB viability, and the relative $TNF-{\alpha}$ expression level (p < 0.05). Moreover, after challenge with Streptococcus iniae ($1{\times}10^8CFU/ml$), the synbiotically fed group exhibited significantly higher (p < 0.05) protection against streptococcosis, validating the observed changes in immune parameters and induction of the cytokine-encoding gene. Therefore, according to the results of the present study, synbiotic feed ($LI_2+BGO$) increased growth, modulated innate immune parameters and protected olive flounder against streptococcosis.