• Journal of Life Science
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• v.26 no.1
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• pp.17-22
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• 2016

Ethanol is a very valuable material, however, it is also a source of stress, as the accumulation of ethanol in a medium inhibits cell viability and decreases productivity of the target product. Therefore, the ethanol tolerance of yeast, which is closely related to ethanol productivity, is an important factor in industrial ethanol production. In this study, the YDJ1 and PEP5 genes were selected as target genes for elucidating ethanol-tolerant mechanisms by analyzing the expression regulation of these genes. The pA-YDJ1 and pA-PEP5 plasmids containing YDJ1 and PEP5 genes under an ADH1 promoter, respectively, were constructed and transformed into BY4742 (host strain), BY4742△ydj1, and BY4742△pep5 strains. The ethanol tolerance in the BY4742△ydj1/ pA-YDJ1 and BY4742△pep5/pA-PEP5 transformants was restored by overexpression of the YDJ1 and PEP5 genes to the host strain level. The YDJ1 and PEP5 genes were also introduced into the double gene disruptant (BY4742△ydj1△pep5) to investigate the expression regulation of the YDJ1 and PEP5 genes. The simultaneous overexpression of the YDJ1 and PEP5 genes restored ethanol tolerance to the 90% level of the BY4742 strain under 8% ethanol stress. The YDJ1 gene induced more overexpression of the PEP5 gene in the BY4742△ydj1 △pep5/pA-YDJ1, pA-PEP5 strain, suggesting that the YDJ1 gene partially regulates the expression of the PEP5 gene as an upstream regulator.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.371-377
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• 2023

In this study, a pepA gene encoding glutamyl (aspartyl)-specific aminopeptidase (PepA; E.C. 3.4.11.7) was cloned from Tetragenococcus halophilus CY54. The translated PepA from T. halophilus CY54 showed very low similarities with PepAs from Lactobacillus and Lactococcus genera. The pepA from T. halophilus CY54 was overexpressed in E. coli BL21(DE3) using pET26b(+). The recombinant PepA was purified by using an Ni- NTA column. The size of the recombinant PepA was 39.13 kDa as determined by SDS-PAGE, while its optimum pH and temperature were pH 5.0 and 60℃, respectively. In addition, the PepA was completely inactivated by 1 mM EDTA, indicating its metallopeptidase nature. The Km and Vmax of the PepA were 0.98 ± 0.006 mM and 0.1 ± 0.002 mM/min, respectively, when Glu-pNA was used as the substrate. This is the first report on PepA from Tetragenococcus species.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.51-56
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• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.155-160
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• 2006

A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.69.1-69
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• 2003

RSH (relA/spoT homolog) has been known to determine the level of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effector nucleotide of the prokaryotic stringent response and also play a role in antibiotic production and differentiation in Streptomyces species but not a little in eukaryotic organism, especially in plant. Salicylic acid (SA), a critical signal molecule of establishing systemic acquired resistance (SAR), could induce SAR in Pepper (Capcicum annuum) against Phytophthora capsici. And the extent of SAR induction was in proportion to the dosage of SA (or BTH). Suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, was carried out between SA-treated and non-SA-treated pepper leaves to isolate genes which may be responsible for defense signaling against pathogens. Early upregulated gene was selected from reverse northern and kinetics of SSH-genes transcripts in SA-treated pepper leaves upon SA treatment. Full-length cDNA of the gene (PepRSH； Pepper RelA / SpoT homolog) had an open reading frame (ORF) of 2166 bp encoding a protein of 722 amino acids and a significant homology with (p)ppGpp phosphohydrolase or synthetase. Genomic DNA gel blot analysis showed that pepper genome has at least single copy of PepRSH. PepRSH transcripts was very low in untreated pepper leaves but strongly induced by SA and methyljasmonic acid (MeJA), indicating that PepRSH may share common SA and MeJA-mediated signal transduction pathway Functional analysis in E. coli showed PepRSH confers phenotypes associated with (p)ppGpp synthesis through a complementation using active site mutagenesis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.3
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• pp.684-692
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• 2003

The herbal extract(YMT_02) is a modified extracts from Yukmijihwang-tang(YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as replenishing yin and tonifying the kidneys herbal medicine for hundred years ia Asian countries. The purpose of this study is to: 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by passive avoidance test, 2) statistical evaluation of candidate gene expression (pentraxin. PEP-19, transthyretin) in rat hippocampus. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Real Time PCR. The results were as follows : 1) passive avoidance test showed enhancing memory retentin by YMT_02 treatment, 2) expression of pentraxin, that accelerate degenerating of neuronal cell, was significantly decreased, 3) the mRNA of genes that has been known to be associated with protecting neuronal cell degeneration, such as PEP-19 and transthyretin, were significantly increased upon YMT_02 treatment. From above results, the administration of YMT_02 which tonify the function of Kidneys could enhance the ability of memory and learning. In addition, the administration of YMT_02 enhance memory retention through modulating particular gene (pentraxin, PEP-19, transthyretin) expressions in hippocampu.

• Journal of Genetic Medicine
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• v.2 no.1
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• pp.35-39
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• 1998

Duchenne/Becker muscular dystrophy are the major neuromuscular disorders with X-linked recessive inheritance. Preimplantation diagnosis of sex determination has been generally used to avoid male pregnancies with these diseases. However, in order to determine if the embryo is normal, carrier or affected regardless of the sex, there is a need for a combined analysis of specific exon on dystrophin gene as well as sex determination of embryo using the same biopsied blastomere. If the exon deletion is not determinable, further diagnosis of carrier or patient can be performed by haplotype analysis. In this study, we applied the primer extension preamplification (PEP) method, which amplifies the whole genome, in 40 cases of single amniocyte and 40 cases of chorionic villus cell. We analysed haplotypes using two (CA)n dinucleotide polymorphic markers located at the end of 5' and 3' region of the dystrophin gene. Exon 46 of dystrophin gene and DYZ3 on chromosome Y were chosen as a target sequence for coamplification PCR. Upon optimizing the conditions, the amplification rates were 91.25% (73/80) for haplotypes (92.5% in amniocyte, 90% in chorionic villus cell) and 88.75% (71/80) for coamplification (85% in amniocyte, 92.5% in chorionic villus cell). The result of the study indicates that haplotypes analysis and coamplification of dystrophin and Y-specific gene using PEP can be applied to prenatal and preimplantation diagnosis in Duchenne/Becker muscular dystrophy making it possible to determine if the fetus is a carrier or an affected one.

• The Plant Pathology Journal
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• v.20 no.4
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• pp.258-263
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• 2004

Ripe fruit of pepper (Capsicum annuum) showed resistance to Colletotrichum gloeoporioides, but unripe fruit was susceptible. We previously isolated the PepTLP gene that induced in both unripe and ripe fruit by fungal infection and wound, and only in ripe fruit by jasmonic acid (JA) treatment. To examine further regulation of PepTLP, the action of specific agonist and antagonists of known signaling effector on the .PepTLP expression by fungal infection, wound, and JA was investigated. A similar dephosphorylation event negatively activated all the PepTLP expression in the ripe fruit by fungal infection, wound, and JA. The induction of PepTLP expression by wound is differentially regulated via phosphorylation and dephosphorylation step during pre- and post-ripening, respectively. In addition, the induction of PepTLP expression in the ripe fruit by wound and JA is differentially regulated via dephosphorylation and phosphorylation step, respectively. Only both wound and JA treatment has synergistic effect on the PepTLP expression in the unripe fruit. Both SA and JA treatments on the unripe fruit, and both wound or JA and SA on the ripe fruit could not do any effect on the expression of PepTLP. These results suggest that the induction of PepTLP expression is differentially regulated via complex regulatory system against fungal infection, wound, and JA treatment during pre- and post-ripening of pepper fruit.

• Bulletin of the Korean Chemical Society
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• v.33 no.2
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• pp.583-588
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• 2012

Functional interaction between Drosophila orphan receptor FTZ-F1 (NR5A3) and a segmentation gene product fushi tarazu (FTZ) is crucial for regulating genes related to define the identities of alternate segmental regions in the Drosophila embryo. FTZ binding to the ligand-binding domain (LBD) of FTZ-F1 is of essence in activating its transcription process. We determined solution structures of the cofactor peptide ($FTZ^{PEP}$) derived from FTZ by NMR spectroscopy. The cofactor peptide showed a nascent helical conformation in aqueous solution, however, the helicity was increased in the presence of TFE. Furthermore, $FTZ^{PEP}$ formed ${\alpha}$-helical conformation upon FTZ-F1 binding, which provides a receptor bound structure of $FTZ^{PEP}$. The solution structure of $FTZ^{PEP}$ in the presence of FTZ-F1 displays a long stretch of the ${\alpha}$-helix with a bend in the middle of helix.

• BMB Reports
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• v.39 no.5
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• pp.642-647
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• 2006

Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.