• Molecules and Cells
• /
• v.24 no.3
• /
• pp.316-322
• /
• 2007

Glutaredoxins (Grxs), also known as thioltransferases (TTases), are thiol oxidoreductases that regulate cellular redox state in a variety of organisms. In the budding yeast Saccharomyces cerevisiae, Grx1 and 2 are cytosolic dithiol Grxs, while Grx3, 4 and 5 are monothiol Grxs. A gene encoding a new monothiol Grx, Grx6, was cloned from the genomic DNA of S. cerevisiae by PCR. Its DNA sequence contains 1,080 bp, and encodes a putative protein of 203 amino acid residues containing Cys-Phe-Tyr-Ser at the active site. Grx6 is similar to other monothiol Grxs in the same organism and to Grx3 in the fission yeast Schizosaccharomyces pombe. and its predicted three-dimensional structure resembles that of S. pombe Grx3. S. pombe cells harboring plasmid pFGRX6 containing the Grx6 gene had about 1.3-fold elevated Grx activity in the exponential phase, and grew better than the control cells under some stressful conditions. Synthesis of ${\beta}$-galactosidase from a Grx6-lacZ fusion gene in S. pombe was enhanced by potassium chloride, aluminum chloride and heat ($37^{\circ}C$) treatment. S. pombe cells harboring plasmid pFGRX6 had elevated ROS levels whereas S. pombe cells harboring extra copies of Grx3 had reduced ROS levels.

• Molecular & Cellular Toxicology
• /
• v.1 no.1
• /
• pp.62-71
• /
• 2005

For toxicity model in the kidney, renal cell carcinoma (RCC) is one of the most important model to assess the structural and functional alterations. Most RCCs are sporadic, and environmental agents are suspected to play a role in the etiology of the disease. In this study, we discovered novel evidence for previously unknown gene expression patterns related to progression according to cancer stage in RCC. Four clear cell RCC tissue samples along with five corresponding patient-matched normal kidney tissue samples were obtained from patients undergoing partial or radical nephrectomy. To examine the difference of gene expression profile in clear cell RCC, radioactive cDNA microarrays were used to evaluate changes in the expression of 1,152 genes in a total. Using $^{33}P-labeled$ probes, this method provided highly sensitive gene expression profiles including drug metabolism, and cellular signaling. 29 genes were identified with expression levels that differed by more than 2.0 value of z-ratio, compared with that in control. Whereas expression of 38 genes were decreased by less than-2.0 value of z-ratio. In conclusion, this study has identified 67 gene expression alterations in clear-cell type of RCC. Most notably, genes involved in cell growth were up-regulated in stage I more than stage III whereas genes involved in signal transduction were down-regulated in which both stage I and stage III. The identified alteraions of gene expression will likely give in sight in to clear cell RCC and tumor progression.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.3
• /
• pp.450-456
• /
• 2006

Pseudomonas fluorescens MC07 is a growth-promoting rhizobacterium that suppresses mycelial growth in fungi such as Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici. To determine the role of the bacterium's antifungal activity in disease suppression, we screened 2,500 colonies generated by Tn5lacZ insertions, and isolated a mutant 157 that had lost antifungal activity. The EcoRI fragment carrying Tn5lacZ was cloned into pBluescript II SK(+) and used as a probe to isolate wild-type clones from a genomic library of the parent strain, MC07. Two overlapping cosmid clones, pEH4 and pEH5, that had hybridized with the mutant clone were isolated. pEH4 conferred antifungal activity to the heterologous host P.fluorescens strain 1855.344, whereas pEH5 did not. Through transposon mutagenesis of pEH4 and complementation analyses, we delineated the 14.7-kb DNA region that is responsible for the biosynthesis of an antifungal compound. DNA sequence analysis of the region identified 11 possible open reading frames (ORF), ORF1 through ORF11. A BLAST search of each putative protein implied that the proteins may be involved in an antifungal activity similar to polyketides.

• Korean Journal of Ecology and Environment
• /
• v.38 no.4 s.114
• /
• pp.510-517
• /
• 2005

The filamentous cyanobacterium Anabaena is capable of both photosynthesis and nitrogen fixation which probably facilitated its incredible adaptation and proliferation in freshwater environments. A small gene, patS, was found to block nitrogen fixing cells from developing which resulted in death of Anabaena in the absence of combined nitrogen sources. We analyzed the DNA sequences in the vicinity of the patS gene by using a codon usage program and detected no codon bias other than the patS open reading frame. Three overlapping cosmids that contain the patS gene were identified, and the presence of other known heterocyst-controlling genes was examined. The patS expression in response to nitrogen starvation was analyzed at the level of transcription and translation by using Northern blot analyses and lacZ-reporter-gene fusion experiments, respectively. The patS expression increased rapidly (within 12 hours) upon the removal of combined nitrogen from the media.

• Korean Journal of Ecology and Environment
• /
• v.44 no.4
• /
• pp.364-372
• /
• 2011

The pale chub (Zacco platypus) is generally found in Asian countries, such as Korea, Japan, and China. Nevertheless, very little information exists about the genes involved in the metabolism of xenobiotics in this species. This species is useful in monitoring the environmental impact on various pollutants in freshwater as a sentinel fish species. We cloned the full-length cDNA sequence of xenobiotic metabolizing cytochrome P450 1A (CYP1A) gene from Z. platypus and characterized it. Tissue distribution and timedependent induction of CYP1A were studied by real-time RT-PCR. Induction pattern of CYP1A was studied by exposing the fish to an arylhydrocarbon receptor agonist, ${\beta}$-naphthoflavone (BNF). The liver showed the highest level of expression in basal state as well as BNF- treated fish. However, appreciable levels of expression were also recorded in Gill and kidney and the least level of expression was observed in the eye. The results of the time-course study revealed an induction in the liver, brain, and gills after 6 h and 12 h in most of the tissues. This study provides an insight into the xenobiotics metabolizing system of Z. platypus and offers baseline information for further research related to biomarker, stress, and adaptive response of this ecologically important fish species in the freshwater environment.

• Microbiology and Biotechnology Letters
• /
• v.16 no.5
• /
• pp.363-368
• /
• 1988

Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

• The Korean Journal of Mycology
• /
• v.22 no.3
• /
• pp.247-253
• /
• 1994

This study was carried out to isolate and characterize $A{\alpha}$ mating locus controlling fruiting body formation directly in the Basidiomycete Schzophyllum commune growing in the North America. Total numbers of genomic library of S. commune UVM1-34 was about $2{\times}10^4$ cells. About 90% library was appeared to have about 35 kb inserted genome DNA in cosmid pTC20 vector. 6 clones were proved to have positive signal to probes within Z and Y region in colony and southern hybridization. In the mating activity test, all the 6 positive clones were appeared to have $A{\alpha}3$ mating activity although they had two different restriction patterns. pSC13 containing 5.7 Kb PstI-fragment of UVM 1-34 $A{\alpha}3$ allele showed about 50% clamp cell formation indicating mating activity when cotransformation was done together with cosmid pTC20.

• The Korean Journal of Mycology
• /
• v.47 no.1
• /
• pp.83-88
• /
• 2019

A previous record of Coleosporium phellodendri on Zanthoxylum ailanthoides in Korea by Korean researchers was regarded as C. zanthoxyli by foreign scientists without further studies. To clarify the identity of a Coleosporium species occurring on Z. ailanthoides in Korea, we examined the morphological characteristics of eight materials and analyzed the internal transcribed spacer (ITS) and 28S large subunit (LSU) rDNA regions of three representative specimens which were collected in Korea and deposited in the Mycological Herbarium at Korea University (Seoul, Korea). All specimens collected in Korea were morphologically and molecularly determined to be C. zanthoxyli. Therefore, this is the first confirmed report on the presence of C. zanthoxyli on Z. ailanthoides in Korea.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.12
• /
• pp.1633-1639
• /
• 2009

This study investigated the effect of maternal undernutrition during late pregnancy on the growth and development of ovine fetal visceral organs. One hundred Mongolian ewes were mated at a synchronized oestrus and divided into three groups and offered 0.175 MJ ME $kgw^{-0.75}\;d^{-1}$ (Restricted Group1; RG1), 0.33 MJ ME $kgw^{-0.75}\;d^{-1}$ (Restricted Group2; RG2) and ad libitum access to feed (Control Group; CG) during late pregnancy (90 days). Selected animals in each group were slaughtered immediately at d 90 of pregnancy and after parturition (neonatal lambs), and major visceral organs were removed and weighed separately. The results indicated that the weights of lung (p<0.01), spleen (p<0.01), heart (p<0.05), liver (p<0.05) and abomasum (p<0.01) in RG1 were significantly lighter than those of CG. For RG2, only the weights of the lung (p<0.05) and spleen (p<0.01) were significantly lighter than those of CG; when expressed as a percentage of body weight, significance was retained in the spleen (p<0.01) for both restricted groups, but the percentage of brain in RG1 was significantly higher than that in CG (p<0.01). For lung and spleen, the amount of DNA was significantly lower (p<0.01) in both groups of restricted neonatal lambs compared to CG; however, there was a significant difference only between RG1 and CG for protein: DNA ratio (p<0.01). The DNA content of kidney, abomasum and jejunum were decreased (p<0.05) in RG1 neonatal lambs, but protein: DNA ratio in the liver was decreased compared with that of CG (p<0.05). The plane of maternal undernutrition during late pregnancy had a significant effect on the growth and development of fetal visceral organs, which altered ontogeny of fetal organ growth and development. These perturbations in fetal visceral development may have significant implications on postnatal growth and adult health.

• Molecules and Cells
• /
• v.27 no.5
• /
• pp.577-582
• /
• 2009

The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ${\gamma}$, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.