• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2000.11a
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• pp.207-208
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• 2000

Since trichloroethylene (TCE), dichloroethylene (DCE), and vinyl chloride (VC) arise from anaerobic degradation of tetrachloroethylene (PCE) and TCE, there is interest in creating aerobic remediation systems that avoid the highly toxic VC and cis-DCE which predonominate in anaerobic degradation. However, it seemed TCE could not be degraded aerobically without an inducing compound (which also competitively inhibits TCE degradation). It has been shown that TCE induces expression of both the toluene dioxygenase of p. putida F1 as well as toluene-p-monooxygenase of P.mendocina KRI. We investigated here the ability of PCE, TCE, and chlorinated phenols to induce toluene-o-xylene monooxygenase (ToMO) from P.stutzeri OX1. ToMO has a relaxed regio-specificity since it hydroxylates toluene in the ortho, meta, and para positions; it also has a broad substrate range as it oxidizes o-xylene, m-xylene, p-xylene, toluene, benzene, ethylbenzene, styrene, and naphthalene; chlorinated compounds including TCE, 1, 1-DCE, cis-DCE, trans-DCE, VC, and chloroform : as well as mixtures of chlorinated aliphatics (Pseudomonas 1999 Maui Meeting). ToMO is a multicomponent enzyme with greatest similarity to the aromatic monooxygenases of Burkholderia pickettii PKO1 and P.mendocina KR1. Using P.sturzeri OX1, it was found that PCE induces P.mendocina KR1 Using P.situtzeri OX1, it was found that PCE induces ToMO activity measured as naphthalene oxygenase activity 2.5-fold, TCE induces 2.3-fold, and toluene induces 3.0 fold. With the mutant P.stutzeri M1 which does not express ToMO, it was also found there was no naphthalene oxygenate activity induced by PCE and TCE; hence, PCE and TCE induce the tow path. Using P.putida PaW340(pPP4062, pFP3028) which has the tow promoter fused to the reporter catechol-2, 3-dioxygenase and the regulator gene touR, it was determined that the tow promoter was induced 5.7-, 7.1-, and 5.2-fold for 2-, 3-, 4-chlorophenol, respectively (cf. 8.9-fold induction with o-cresol) : however, TCE and PCE did not directly induce the tou path. Gas chromatography and chloride ion analysis also showed that TCE induced ToMO expression in P.stutzeri OX1 and was degraded and mineralized. This is the first report of significant PCE induction of any enzyme as well as the first report of chlorinated compound induction of the tou operon. The results indicate TCE and chlorinated phenols can be degraded by P.stutzeri OX1 without a separate inducer of the tou pathway and without competitive inhibition.

• Journal of Life Science
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• v.21 no.6
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• pp.893-900
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• 2011

Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-${\kappa}B$, TNF-${\alpha}$, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

• Journal of Life Science
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• v.14 no.1
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• pp.154-162
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• 2004

Thrombospondin-1 (TSP-1), a negative regulator in tumor growth and angiogenesis, is cell-type specifically regulated and at transcriptional level by external stimuli. Previously, we found that phorbol 12-myristate 13-acetate (PMA) suppressed TSP-1 expression in porcine aortic endothelial (PAE) cell, but enhanced in hepatoma cell line, Hep 3B cell. A region between -767 and -723 on the tsp-1 promoter was defined as a responsive site to the suppression in PAE cell. eased on the previous results, the molecular mechanism of TSP-1 expression was determined by characterizing interactions between cis-elements and trans-factors using three overlapped oligonucleotide probes, oligo a-1 (from -767 to -738), a-2 (-759 to -730) and a-3 (-752 to -723). The results from electromobility shift assay showed that PMA-induced suppression of TSP-1 transcription in PAE cell might be caused via a negative regulator binding to the region from -752 to -730 and additionally generated by lacking two positive regulators binding to the sites from -767 to -760 and from -752 to -730. Especially, PMA enhanced the binding ability of the negative regulator to the site from -752 to -730 in PAE cell, but anti-c-Jun did not affected its binding ability.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.102-109
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• 2010

Fructan has been found to accumulate in various tissues during periods when light levels increased carbon fixation where low temperatures reduced growth rates while photosynthesis continued. In this study, we have cloned 1-sucrose:sucrose fructosyl transferase(1-sst) and 1-fructan: fructan fructosyl transferase (1-fft, a key enzyme for the synthesis of fuctan) from Jerusalem Artichoke (Helianthus tuberosus L.). The recombinant vector with 1-sst and 1-fft has been constructed under the control of 35S promoter of KJGV-B2 vector and transgenic plants obtained by Agrobacterium tumefaciens LBA4404. PCR analysis carried out on the putative transgenic plants for amplification of the coding region of specific gene (1-sst, 1-fft), and HPT genes. Transgenic lines carrying of 1-sst and 1-fft were confirmed for integration into the rice genome using Southern blot hybridization and RT-PCR. The transgenic plants in $T_2$ generation were selected and expression pattern analysis revealed that 1-sst and 1-fft were stable. This analysis confirmed the presence of low-molecular-weight fructan in the seedling of the transgenic rices. Therefore, cold tolerance and carbohydrate metabolism will be possible to develop resistant plants using the transgenic rice.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.145-152
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• 2007

Transgenic mice were generated by microinjecting a plasmid DNA containing the SV40 (simian virus 40) large T antigen (Tag) gene fused with mouse albumin promoter/enhancer sequences into fertilized one-cell mouse embryos. Among eleven founder transgenic animals, four developed hepatocellular carcinoma, two showed kidney cancer and one developed skin and brain tumors. Three stable transgenic lines, #1-2, #1-6 and #1-11 were established. Members of the lines #1-6 and #1-11 reproducibly developed liver tumors by 8 to 10 weeks of age but did not exhibit any phenotypic changes in other tissues. Histological changes loading to liver tumor formation occurred with predictable kinetics and could be classified into three distinct stages; (a) newborn to 3 weeks of age, characterized by hyperplastic hepatocytes with reduced amounts of cytoplasm without any nuclear alterations, (b) between 4 to 8 weeks of age, characterized by diffuse liver cell dysplasia without observable tumor nodules, and (c) 9 weeks of age and thereafter, characterized by hepatocellular carcinomas in the background of extensive liver dysplasia. Metastasis to the lung from a liver carcinoma was observed in #1-11 founder animal. This transgenic mouse system displays similarities with human liver cancers in a number of aspects and provides a useful model for the study of molecular events involved in hepatocarcinogenesis.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.187-191
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• 2007

This study was canted out to develop cell lines overexpressing human H-transferase (HT). One of the approaches to prevent hyperacute rejection in xenotransplantation might be the expression of human HT in porcine cells. In this study, we cloned human HT gene from HepG2 cells using RT-PCR to establish HT-overexpressing vector. The full-length cDNA of human HT was inserted into the 3' end of CMV promoter for construction of the overexpression vector pRc/CMV-hHT. Using ietPEI DNA transfection reagent, the vector was introduced into porcine ear skin fibroblasts from newborn piglets. Transfected cells were selected by treatment of $300{\mu}g/ml$ G418 for 12 days. After antibiotic selection, survived colonies with approximately 5mm in diameter were picked and analysed for transgene human HT by PCR. The colonies proven to be human HT transfectants were analysed by RT-PCR to determine their expressions or human HT. In all colonies tested, human HT mRNA was detected. This result demonstrates the establishment of porcine cell lines overexpressing human HT, and these cell lines may be used for the development of transgenic pigs for xenotransplantation.

• Journal of Gastric Cancer
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• v.21 no.4
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• pp.379-391
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• 2021

Purpose: Promoter DNA methylation of various genes has been associated with metachronous gastric cancer (MGC). The cancer-specific methylation gene, cysteine dioxygenase type 1 (CDO1), has been implicated in the occurrence of residual gastric cancer. We evaluated whether DNA methylation of CDO1 could be a predictive biomarker of MGC using specimens of MGC developing on scars after endoscopic submucosal dissection (ESD). Materials and Methods: CDO1 methylation values (TaqMeth values) were compared between 33 patients with early gastric cancer (EGC) with no confirmed metachronous lesions at >3 years after ESD (non-MGC: nMGC group) and 11 patients with MGC developing on scars after ESD (MGCSE groups: EGC at the first ESD [MGCSE-1 group], EGC at the second ESD for treating MGC developing on scars after ESD [MGCSE-2 group]). Each EGC specimen was measured at five locations (at tumor [T] and the 4-point tumor-adjacent noncancerous mucosa [TAM]). Results: In the nMGC group, the TaqMeth values for T were significantly higher than that for TAM (P=0.0006). In the MGCSE groups, TAM (MGCSE-1) exhibited significantly higher TaqMeth values than TAM (nMGC) (P<0.0001) and TAM (MGCSE-2) (P=0.0041), suggesting that TAM (MGCSE-1) exhibited CDO1 hypermethylation similar to T (P=0.3638). The area under the curve for discriminating the highest TaqMeth value of TAM (MGCSE-1) from that of TAM (nMGC) was 0.81, and using the cut-off value of 43.4, CDO1 hypermethylation effectively enriched the MGCSE groups (P<0.0001). Conclusions: CDO1 hypermethylation has been implicated in the occurrence of MGC, suggesting its potential as a promising MGC predictor.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.52-69
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• 2022

Ferulic acid (FA) is a phytochemical with various bioactive properties. It has recently been proposed that due to its phytogenic action it can be used as an alternative growth promoter additive to synthetic compounds. The objective of the present study was to evaluate the growth performance, carcass traits, fiber characterization and skeletal muscle gene expression on hair-lambs supplemented with two doses of FA. Thirty-two male lambs (n = 8 per treatment) were individually housed during a 32 d feeding trial to evaluate the effect of FA (300 and 600 mg d^-1) or zilpaterol hydrochloride (ZH; 6 mg d^-1) on growth performance, and then slaughtered to evaluate the effects on carcass traits, and muscle fibers morphometry from Longissimus thoracis (LT) and mRNA abundance of β₂-adrenergic receptor (β₂-AR), MHC-I, MHC-IIX and IGF-I genes. FA increased final weight and average daily gain with respect to non-supplemented animals (p < 0.05). The ZH supplementation increased LT muscle area, with respect to FA doses and control (p < 0.05). Cross-sectional area (CSA) of oxidative fibers was larger with FA doses and ZH (p < 0.05). Feeding ZH increased mRNA abundance for β₂-AR compared to FA and control (p < 0.05), and expression of MHC-I was affected by FA doses and ZH (p < 0.05). Overall, FA supplementation of male hair lambs enhanced productive variables due to skeletal muscle hypertrophy caused by MHC-I up-regulation. Results suggest that FA has the potential like a growth promoter in lambs.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.1-35
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• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

• 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.