• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.189-189
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• 2022

Soil salinity negatively affects plant growth, productivity, and metabolism. Rice is known to have more sensitive phenotypes than other cereal crops, such as wheat, sorghum, and barley. We characterized the molecular function of rice C3HC4 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under normal conditions, there is no significant phenotypic difference between transgenic plants and WT plants. However, OsRFPH2-3-overexpressing plants exhibited higher fresh weight and length under saline conditions. Also, transgenic plants maintain higher chlorophyll, proline, and soluble sugar contents and lower H₂O₂ and MDA contents than the wild type; these results support transgenic plants with enhanced salinity tolerance phenotypes.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.263-269
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• 2008

Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

• BMB Reports
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• v.33 no.6
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.11
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• pp.4465-4466
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• 2014

HPV type-specific detection may promote cervical screening program and vaccination development worldwide. We conduct a study comparing HPV Hybrid capture II (HC II) Test and Hybribio GenoArray test, a newly developed HPV type-specific assay, in patients with cervical epithelial neoplasm. Results showed a good concordance in cervical HPV detection between two tests (kappa value 0.80, p<0.05, McNemar test). Our study may promote utilization of type-specific HPV detection that is helpful for cervical cancer screening and vaccination.

• Korean Journal of Pharmacognosy
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• v.40 no.3
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• pp.196-204
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• 2009

This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.239-255
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• 1997

Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

• Journal of Animal Science and Technology
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• v.53 no.2
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• pp.113-118
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• 2011

This study was conducted to evaluate effects of enzyme complex on growth performance, nutrient digestibility, blood profiles and feed cost in growing pigs. Ninety-six pigs [(Landrace ${\times}$ Yorkshire) ${\times}$ Duroc, $22.96{\pm}0.79$ kg average initial body weight] were used in 42d growth assay. Dietary treatments included:1) HC (high energy and nutrient density diet), 2) CON (control, basal diet), 3) CE1 (CON + 0.05% enzyme complex) and 4) CE2 (CON + 0.1% enzyme complex). Four pigs were allotted per pen with six replicate pens per treatment by completely randomized design. The ADG was higher in CE1 and CE2 treatments than CON treatment (P<0.05). The ADFI was linearly increased by CE treatments compared to HC treatment. The CE1 treatment had highest DM, N and GE digestibility (P<0.05). Digestibility of DM, N and GE were quadratic enhanced by enzyme complex level. No differences were found among treatments for creatinine and BUN. The enzyme complex treatments (CE1 and CE2) showed lower feed cost/body weight gain than HC treatment. In conclusion, enzyme complex can improve ADG and reduce feed cost/body weight gain when low energy diet was used. Furthermore adding 0.05% enzyme complex had highest nutrient digestibility.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.90-95
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• 2001

Kinetic Analysis of Cathepsin B Inhibitor Using a Spectrophotometric Assay. Han, Kil-Hwan and SangDal Kim*. Department of Applied MicrobioJ0f5Yt Yeungnam UniversitYt Kyongsan 77 2-749, Korea - The KHS 10, C4Hl10~6 formula produced from Streptomyces luteogriseus KT-] 0 effectively inhibited a lysosomal cysteine proteinase, cathepsin B. It inhibited the enzyme activity of cathepsin B competitively when the N a-CBZ-Llysine p-nitrophenyl ester HC] (CLN) was used as a substrate. The inhibition const:mt (Ki) of KHS 1 0 for cathepsin B detennined by spectrophotometeric assay was 430 nM. The effective inhibition of cathepsin B was observed at $25^{\circ}C$ :md pH 6.0. The cathepsin B inhibitor, KHSlO needed a preincubation of cathepsin B with the inhibitor for over 5 min. The KHS 10 preserved over 80% inhibition activity even after heat-treatment at $100^{\circ}C$ for ] hr.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.88-88
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• 2003

Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

• Korean Journal of Health Education and Promotion
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• v.27 no.1
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• pp.1-8
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• 2010

Objectives: Human papillomavirus (HPV) infection is highly associated with cervical cancer. So, the modification of the risk factors of HPV infection is essential for prevention of cervical cancer. This study was performed to evaluate the risk factors of HPV infection. Methods: HPV test of 12,337 study population conducted using Hybrid-Capture II assay(HC-II) and self-administered questionnaires were collected. The study population was people who visited hospital-based medical screening center from January to December 2007 and all were female employees or employees' partner. Results: In logistic regression analysis, smoking and alcohol drinking were significant factors, with odds ratios of 1.328 (95% CI 1.010~1.746) and 1.644 (95% CI 1.309~2.066), respectively. Nutritional supplements was also significant factor, which odds ratio was 1.161 (95% CI 1.004~1.343). Oral contraceptives was positive association with HPV infection (odds ratio 2.108; 95% CI 1.217~3.652), whereas condom was negative association (odds ratio 0.851; 95% CI 0.740~0.979). Conclusion: HPV Prevalence of 12,377 study population was 11.4%. Smoking, alcohol drinking, nutritional supplements and oral contraceptives were possible risk factors of HPV infection, and condom had possible preventive effect on HPV infection. Further prospective and comprehensive studies about HPV risk factors are required.