• BMB Reports
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• v.39 no.5
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• pp.595-606
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• 2006

Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.

• The Journal of Natural Sciences
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• v.9 no.1
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• pp.31-37
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• 1997

Apoptosis is an important event in the anticancer drug therapy. p53 was demonstrated to serve a key component to lead tumor cell death by inducing apoptosis. However, recent study showed the presence of p53 independent apoptotic pathway (Gaftenhaus et al., 1996). We were curious to know it apoptosis induced by adriamycin, a genotoxic anticancer agent, involved p53 gene mutation. Thus this study investigated the p53 gene mutation status among HeLa cell population during apoptosis induced by adriamycin. Under our experimental condition, 12 hour treatment of 1 ${\mu}m$ adriamycin caused apoptosis which was monitored by DNA fragmentation assay. In order to see the p53 gene mutation status, exons of 5, 7 and 8 of p53 gene, where previously reported p53 mutation hot spots reside, were amplified by PCR and nucleotide sequence change was scanned. However, no nucleotide change was observed among apoptotic HeLa cell population. Therefore this study demonstrated that adriamycin induced apoptosis without causing p53 gene damage.

• Journal of Animal Science and Technology
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• v.53 no.6
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• pp.511-516
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• 2011

Fas gene known to associate with intramuscular fat content in Korean cattle was selected for DNA marker development. Fas (APO-1, CD95), a member of the tumor necrosis factor (TNF) receptor superfamily, is a cell membrane protein that mediates apoptosis (programmed cell death). We discovered single nucleotide polymorphisms (SNPs) within Fas gene in order to develop novel DNA markers at genomic level. Of this gene to search for SNP, sequences of whole exon and 1kb range of both front and back of the gene using 24 cattle were determined by direct-sequencing methods. As a result, 16 SNPs in exon, 37 SNPs in intron and 2 SNPs in promoter region, a total of 55 SNPs were discovered. In these SNPs, thirty-one common polymorphic sites were selected considering their allele frequencies, haplotype-tagging status and Linkage Disequilibrium (LD) for genotyping in larger-scale subjects. Selected SNPs were confirmed genotype through SNaPshot method (n=274) and were examined for possible genetic association of Fas polymorphisms with carcass weight (CWT), eye muscle area (EMA), and backfat thickness (BF). So, the SNP have been identified significant g.-12T>G, g.1112T>G and g.32548T>C. These results suggest that polymorphism of Fas gene was associated with meat quality traits in Hanwoo.

• Korean Journal of Plant Tissue Culture
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• v.28 no.5
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• pp.255-261
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• 2001

Vitis vinifera thaumatin-like protein (VVTL1) is a fruit-specific and ripening-related protein in grape. In order to isolate VVTL1-homolog gene and fruit-specific promoter from Campbell cultivar, we isolated a genomic clone containing VVTL1-homolog gene from grape genomic library through plaque hybridization. VVTL1-homolog gene has an intronless genomic structure, which the pattern is matched with those of other PR5 genes such as osmotin and osmotin-like protein genes. Transcription start site was determined by primer extension analysis. The promoter region of VVTL1-homolog gene contains a sequence or structure, especially the location and number of TCA box and ABRE (abscisic acid-responsive element), distinct from other reported plant PR5 genes, though with several known functional elements such as a TATA box and CAAT box. These results suggested that VVTL1-homolog gene may be regulated by a plant hormone, abscisic acid, and one or several stresses such osmotic pressure and pathogen infection. The isolation of fruit-specific promoter may be helpful to breed a genetically modified grape with valuable phenotype or materials in fruits.

• Korean Journal of Microbiology
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• v.29 no.1
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• pp.16-24
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• 1991

The yeast gene THR1 encodes the homoserine kinase (EC 2.7.1.39: HKase) which catalyses the first step of the threonine specific arm at the end of the common pathway for methionine and threonine biosynthesis. A recombinant plasmid pMC3 (12.6 kilobase pairs, vector YCp50) has been cloned into E. coli HB101 from a yeast genomic library through its complementing activity of a thr1 mutation in a yeast recipient strain M39-1D. When subcloned into pMC32 (8.6kbp, vector YRp7) and pMC35 (8.3 kbp, vector YIp5), the HindIII fragment (2.7 kbp) of pMC3 insery was positive in the thrI complementing activity in both yeast and E. coli auxotrophic strains. The linearized pMC35 was introduced into the original recipient yeast strain and the mitotically stable chromosomal integrant was identified among the transformants. Through the tetrad analysis, the integration site of the pMC35 was localized to the region of THR1 structural gene at an expected genetic distance of approximately 11.1 cM from the ARG4 locus on the right arm of the yeast chromosome VIII. When episomically introduced into the auxotrophic cells and cultured in Thr omission liquid medium, the cloned gene overexpressed the HKase in the order of thirteen to fifteenfold, as compared with a wildtype. HKase levels are repressed by addition of threonine at the amount of 300 mg/l and 1, 190 mg/l for pMC32 and pMC3, respectively. Data from genetic analysis and HKase response thus support that the cloned HindIII yeast DNA fragment contains the yeast thr1 structural gene, along with necessary regulatory components for control of its proper expression.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.3
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• pp.285-292
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• 1997

This study was conducted to obtain the transgenic Brnssica napus plants with tobacco Apase gene using the binary vector system of Agrobacteriurn fumefociens. The results obtained were summarized as follows: A repressible acid phosphatase gene of Saccharon~yces cerevisiae, pho105 was used for screening of tobacco Apase cDNA. In order to identify Apase gene in tobacco genome, Southern blot analysis was pcrformed and the Apase gcnc may be present as a single copy, or at most two or three copies, in tobacco genome. To isolate the tobacco Apase gene, tobacco cDNA library was constructed using purifed mRNA from -Pi treated tobacco root and the plaque forming unit of the library was 2.8 x $10^5$ pfu/m${\ell}$, therefore the library might cover all expressed mRNAs. Using pho5 as a probe. tobacco Apase cDNA was cloned, and restriction mapping and Southern blot analysis of cDNA insert were revealed that the 3.6 kb cDNA contained tobacco acid phosphatase cDNA. Plasmid pGA695 -tcAPl was constructed by subcloning tobacco Apase cDNA into the Hind site of pGA695 with 35s promoter which can be expressed constitutively in plants. The Brassica napus cotyledonary petioles were cocultivated with the ,4 grobacteriunz and transferred to the selection medium. The transformed and regenerated plants were transplanted to soil medium. Southern blot analysis was done on the transformed plants, and it was confirmed that a foregin gene was stably integrated into the genonies of B. nnpus plants.

• Journal of Life Science
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• v.21 no.12
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• pp.1784-1788
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• 2011

The typical miRNA and its nearby host gene are co-expressed by sharing the same promoter. We assumed that miR-7b and its host gene FICT might use an identical promoter for their brain specific gene expression. Sequence comparison of the genomic DNA of mouse miR-7b, human miR-7-3 and their host genes by using the bioinformatic tools revealed high sequence homology and several putative transcription factor-binding sites on the promoter region. In order to probe the hypothesis we used a luciferase vector system into which we cloned the 5' upstream conserved region of miR-7b and FICT. The putative promoter region showed decreased luciferase activity, suggesting that the 5' upstream of miR-7b and FICT contain a negative regulator for gene expression.

• Journal of Life Science
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• v.16 no.1
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• pp.126-130
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• 2006

The present study intends to characterize the DNA damage-inducible responses in yeast. The fission yeast, Schizosaccharomyces pombe was used in this study as a model system for higher eukaryotes. To study UV-inducible responses in S. pombe, five UV-inducible cDNA clones were isolated from S. pombe by using subtration hybridization method. To investigate the expression of isolated genes, UVI-155, the cellular levels of the transcripts were determined by Northern blot analysis after UV-irradiation. The transcripts of isolated gene (UVI-155) increased rapidly and reached maximum accumulation after UV-irradiation. Compared to the message levels of control, the levels of maximal increase were approximately 5 fold to UV-irradiation. In order to investigation whether the increase of UVI-l55 trascripts was a specific results of UV-irradiation, UVI-155 transcript levels were examined after treating the cells to mthylmethane sulfonate (MMS). The transcripts of UVI-155 were not induced by treatment of $0.25\%$ MMS. These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of these genes. To characterize the UVI-155 gene, gene deletion experiments were analyzed. The deleted strain was not well grown. This result indicated that the UVI-155 gene is essential for cell viability.

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.89-94
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• 2004

This study was conducted to determine expression of acyl-CoA synthetase 4(ACS4), which is involved in converts arachidonic acid to postaglandins, in the mouse uterus during pregnancy. In arachidonic acid metabolism, acyl-CoA synthetase plays a key role in the esterification of free arachidonic acid into membrane phospholipids. Following its release by the action of calcium dependent phospholipases, free arachidonic acid is believed to be rapidly converted to arachidonoyl-CoA and reesterified into phospholipids in order to prevent excessive synthesis of prostaglandins. Here we demonstrate that ACS4 gene are differentially regulated in the peri-implatation mouse uterus. During the preimplantation period(days 0.5∼3.5), the ACS4 gene was expressed in the uterus until day 3.5 after which the expression was downregulated. The expression of cPLA2, COX1, and COX2 gene was similar to that of ACS4 gene in the preimplantation periods. However expression levels of COX1 gene show much variation on the various days of pregnancy examined. These data, suggest that ACS4 expression in preimplantation period is involved in initial attachment reaction with cPLA2, COX1, and COX2 gene.

• Korean Journal of Breeding Science
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• v.41 no.3
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• pp.194-204
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• 2009

Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.