• Korean Journal of Exercise Nutrition
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• v.17 no.2
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• pp.49-55
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• 2013

Our purpose was to investigate the effect of a 5-week high-fat diet on the body weight, blood components [triglycerides (TGs), free fatty acids (FFAs), and glucose], and lipoprotein lipase (LPL) mRNA expression in the skeletal muscles of rats. Body weight increased overtime in experimental and control groups without significant differences. In terms of the blood components, the density of TG was significantly lower in the high-fat diet group compared to the control, whereas FFA and glucose levels were similar in the two groups. Although the levels of glyceraldehyde 3-phosphate dehydrogenase(GAPDH) mRNA expression were similar between the groups, LPL mRNA expression was significantly higher in the high-fat diet group than that in the control group. The results of this study suggest that a high-fat diet enhances LPL mRNA expression, as well as possibly increases fat metabolism. For a better understanding of the relationship between diet patterns and fat metabolism, further analysis of genes related to the fat metabolism is warranted.

• KSBB Journal
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• v.10 no.1
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• pp.38-45
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• 1995

The expression of Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpdA) and trpC promoters in A. oryzae were compared using E. coli lacZ gents fusions. The specific activities of the expressed E. coli $\beta$-galactosidase in A. oryzae transformants containing the A. nidulans gpdA promoter were around 2,000 units per ug of protein. The specific activities of transformants containing the A. nidulans trpC promoter were very low, ranging from 10.5 to 52.3 units per ug of protein. These results showed that the expression of the A. nidulans gpdA promoter in A. oryzae was approximately 70 times greater than the A. nidulans trpC promoter. In western blot analysis, immunoreactive bands of a imlilar molecular weight as the E. coli $\beta$-galactosidase were observed in A. oryzae carrying the gpdA-lacZ fusion and to a lesser intensity in those carrying the tvpC-lacZ fusion. Southern analysis showed that the higher expression of the gpdA-lacZ fusion as compared to the trpC-lacZ fusion was not due a greater number of integrated plasmids.

• Parasites, Hosts and Diseases
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• v.58 no.6
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• pp.675-679
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• 2020

MYB2 protein was identified as a transcription factor that showed encystation-induced expression in Giardia lamblia. Although nuclear import is essential for the functioning of a transcription factor, an evident nuclear localization signal (NLS) of G. lamblia MYB2 (GlMYB2) has not been defined. Based on putative GlMYB2 NLSs predicted by 2 programs, a series of plasmids expressing hemagglutinin (HA)-tagged GlMYB2 from the promoter of G. lamblia glutamate dehydrogenase were constructed and transfected into Giardia trophozoites. Immunofluorescence assays using anti-HA antibodies indicated that GlMYB2 amino acid sequence #507-#530 was required for the nuclear localization of GlMYB2, and this sequence was named as NLSGlMYB2. We further verified this finding by demonstrating the nuclear location of a protein obtained by the fusion of NLSGlMYB2 and G. lamblia glyceraldehyde 3-phosphate dehydrogenase, a non-nuclear protein. Our data on GlMYB2 will expand our understanding on NLSs functioning in G. lamblia.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.80.1-80
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• 2003

Several plant and microbial genes that could confer disease resistance in transgenic rice plants are being cloned and characterized. We are currently constructing transgenic rice lines that overexpress the gene products, such as a galactinol synthase, a defensin, and a bacterial ACC deaminase. Subtractive hybridization of a rice cDNA library constructed from the Xanthomonas oryzae-infected ice leaves resulted in isolation of many inducible cDNA clones including a elongation factor EF2, a oryzain alpha, a catalase, a aldehyde dehydrogenase, a S-adenosylmethionine synthetase, a caffeic acid O-methyltransferase, a glyceraldehyde-3-phosphate dehydrogenase, a light-regulated protein, nKY transcription factors, and a nucleotide diphosphate kinase. Some genes among those may be useful genetic sources for construction of disease resistant transgenic rice. Full lengths of the rice OsFIERG and a rice oryzain genomic clones were cloned, and serial deletion fragments of the promoter regions of these genes were fused with GUS reporter gene in pCAMBIA1201, respectively. Promoter activities of these constructs will be examined upon various stresses and Pathogen infections to obtain the pathogen specific inducible-promoter. This work was supported by a grant from BioGreen 21 Program, Rural Development Administration, Republic of Korea.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.199-202
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• 2008

A white rot fungus Irpex lacteus produced lignin degrading enzymes, which showed degrading activity against various recalcitrant compounds. However, laccase, one of the lignin degrading enzymes, was too low to be assayed by spectrophotometry using o-tolidine as the chromogenic substrate in this fungus under various culture conditions. A laccase expression vector was constructed using a cDNA from Phlebia tremellosa with the constitutively expressed promoter of glyceraldehydes-3-phosphate dehydrogenase gene, and introduced into I. lacteus by the restriction enzyme mediated integration transformation through the protoplast-$CaCl_2$ procedure. Two transformants showed highly increased laccase activities at the early growth phase in the minimal liquid medium, and they not only degraded bisphenol A, a notorious endocrine disrupting chemical, but also removed the estrogenic activity effectively.

• Journal of the korean academy of Pediatric Dentistry
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• v.35 no.1
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• pp.30-38
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• 2008

Dental pulp cells are assumed to possess the capacity to elaborate both bone and dentin matrix under the pathological conditions following tooth injury. The purpose of this study is to examine the effects of mineral trioxide aggregate (MTA) on various gene expression regarding dentinogenesis and cell viability assay in cultured primary human dental pulp cells. The author also examined the effects of this material on cellular alkaline phosphatase activity as a potential indicator of dentinogenesis. For gene expression on MTA, reverse transcriptase polymerase chain reaction was performed using primer sets of glyceraldehyde-3-phosphate dehydrogenase, type I collagen, alkaline phosphatase(ALP), osteonectin, and dentin sialoprotein after 2 and 4 days. Cell viability assay showed that the proportion of MTA-treated pulp cells which had been exposed for 5 days to MTA was higher than that of the control cells. Among the genes investigated in this study, ALP and osteonectin(SPARC) were increased in MTA treated group than in control. These findings suggest that this dental pulp culture system may be useful in the future as a model for studying the mechanisms underlying dentin regeneration after the treatment with MTA. Exposure to MTA material would not induce cytotoxic response in the dental pulp cells. In addition, MTA could influence the behavior of human pulp cells by increasing the ALP activity and SPARC synthesis.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.508-513
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• 2010

Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

• Tuberculosis and Respiratory Diseases
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• v.70 no.4
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• pp.301-306
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• 2011

Background: It is well-known that cell-free nucleic acids rise in patients with many types of malignancies. Several recent experimental studies using cancer cell lines have shown that changes in cell-free RNA are predictive of the response to chemotherapy. The objective of this study was to determine whether quantification of free RNA can be used as a biomarker for clinical responses to chemotherapy in patients with lung cancer. Methods: Thirty-two patients with lung cancer (non-small cell lung cancer, n=24; small cell lung cancer, n=8) were divided into 2 groups according to their responses to chemotherapy (response group, n=19; non-response group, n=13). Blood samples were collected before and after two cycles of chemotherapy. Real-time quantitative RT-PCR was used for transcript quantification of the glyceraldehyde-3-phosphate dehydrogenase gene. Results: The pre chemotherapy values (Response group $41.36{\pm}1.72$ vs. Non-response group $41.33{\pm}1.54$, p=0.78) and post chemotherapy values (Response group $39.92{\pm}1.81$ vs. Non-response group $40.41{\pm}1.47$, p=0.40) for cell free RNA concentrations, expressed as Ct GAPDH (threshold cycle glyceraldehyde-3-phosphate dehydrogenase gene) levels, was not different between the two groups. There was no significant relationship between changes in the cell free RNA level clinical responses after chemotherapy (p=0.43). Conclusion: We did not find a correlation between quantification of serum cell free RNA levels and clinical responses to chemotherapy in patients with lung cancer. Further investigations are needed to determine whether the cell free RNA level is a useful predictor of responses to chemotherapy in patients with lung cancer.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.64-70
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• 2019

Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

• The Plant Pathology Journal
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• v.37 no.4
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• pp.329-338
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• 2021

Alternaria leaf blight is one of the most common diseases in watermelon worldwide. In Korea, however, the Alternaria species causing the watermelon leaf blight have not been investigated thoroughly. A total of 16 Alternaria isolates was recovered from diseased watermelon leaves with leaf blight symptoms, which were collected from 14 fields in Korea. Analysis of internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and RNA polymerase II second largest subunit (RPB2) were not competent to differentiate the Alternaria isolates. On the contrary, analysis of amplicon size of the histone H3 (HIS3) gene successfully differentiated the isolates into three Alternaria subgroups, and further sequence analysis of them identified three Alternaria spp. Alternaria tenuissima, A. gaisen, and A. alternata. Representative Alternaria isolates from three species induced dark brown leaf spot lesions on detached watermelon leaves, indicating that A. tenuissima, A. gaisen, and A. alternata are all causal agents of Alternaria leaf blight. Our results indicate that the Alternaria species associated watermelon leaf blight in Korea is more complex than reported previously. This is the first report regarding the population structure of Alternaria species causing watermelon leaf blight in Korea.