• Journal of Life Science
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• v.23 no.1
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• pp.1-7
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• 2013

Understanding salt tolerance mechanisms is important for the increase of crop yields, and so, several screening approaches were developed to identify plant genes which are involved in salt tolerance of plants. Here, we transformed the Arabidopsis cDNA library into a salt-sensitive calcineurin (CaN)-deficient ($cnb{\Delta}$) yeast mutant and isolated the colonies which can suppress salt-sensitive phenotype of $cnb{\Delta}$ mutant. Through this functional complementation screen, a total of 34 colonies functionally suppressed the salt-sensitive phenotype of $cnb{\Delta}$ yeast cells, and sequencing analysis revealed that these are 9 genes, including CaS, AtSUMO1 and AtHB-12. Among these genes, the ectopic expression of CaS gene increased salt tolerance in yeast, and CaS transcript was up-regulated under high salinity conditions. CaS-antisense transgenic plants showed reduced root elongation under 100 mM NaCl treatment compared to the wild type plant, which survived under 150 mM NaCl treatment, whereas CaS-antisense transgenic plant leaves turned yellow under 150 mM NaCl treatment. These results indicate that the expression of CaS gene is important for stress tolerance in yeast and plants.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.403-410
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• 2017

Background: Prenyltransferases catalyze the sequential addition of isopentenyl diphosphate units to allylic prenyl diphosphate acceptors and are classified as either trans-prenyltransferases (TPTs) or cis-prenyltransferases (CPTs). The functions of CPTs have been well characterized in bacteria, yeast, and mammals compared to plants. The characterization of CPTs also has been less studied than TPTs. In the present study, molecular cloning and functional characterization of a CPT from a medicinal plant, Panax ginseng Mayer were addressed. Methods: Gene expression patterns of PgCPT1 were analyzed by quantitative reverse transcription polymerase chain reaction. In planta transformation was generated by floral dipping using Agrobacterium tumefaciens. Yeast transformation was performed by lithium acetate and heat-shock for $rer2{\Delta}$ complementation and yeast-two-hybrid assay. Results: The ginseng genome contains at least one family of three putative CPT genes. PgCPT1 is expressed in all organs, but more predominantly in the leaves. Overexpression of PgCPT1 did not show any plant growth defect, and its protein can complement yeast mutant $rer2{\Delta}$ via possible protein-protein interaction with PgCPTL2. Conclusion: Partial complementation of the yeast dolichol biosynthesis mutant $rer2{\Delta}$ suggested that PgCPT1 is involved in dolichol biosynthesis. Direct protein interaction between PgCPT1 and a human Nogo-B receptor homolog suggests that PgCPT1 requires an accessory component for proper function.

• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.273-278
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• 2004

Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na$^{+}$ sensitivity of yeast mutant (cnb$\Delta$) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.ant salt stress.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.221-224
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• 1989

Amylolytic filamentous fungus, Aspergillus oryzae and nonamylolytic sugar fermentable yeast, Saccharomyces cerevisiae were fused by protoplast fusion in order to develope microorganisms having their intergrated function. Aminoacid auxotrophic properties were used as a genetic marker of protoplast fusion, and 35% PEG 4000 was used as a fusogenic agent. Complementation frequengy of fusion was $4.6\times 10^{-6}$ Obtained fusants showed the morphology of yeast strains, the amylase activity and the ethanol productivity. Among the properties of the fusants, morphology and prototrophic property were sustained stably but their ethanol productivity from starch was reduced. Although fusant strains had 0.5-fold ethanol productivity compared to that of S. cerevisiae in glucose medium, they produced ethanol from strach by direct fermentation.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.102-108
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• 1994

Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

• Molecules and Cells
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• v.26 no.3
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• pp.270-277
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• 2008

This study addresses the physiological functions of the Ran-binding protein homolog NbRanBP1 in Nicotiana benthamiana. Virus-induced gene silencing (VIGS) of NbRanBP1 caused stunted growth, leaf yellowing, and abnormal leaf morphology. The NbRanBP1 gene was constitutively expressed in diverse tissues and an NbRanBP1:GFP fusion protein was primarily localized to the nuclear rim and the cytosol. BiFC analysis revealed in vivo interaction between NbRanBP1 and NbRan1 in the nuclear envelope and the cytosol. Depletion of NbRanBP1 or NbRan1 reduced nuclear accumulation of a NbBTF3:GFP marker protein. In the later stages of development, NbRanBP1 VIGS plants showed stress responses such as reduced mitochondrial membrane potential, excessive production of reactive oxygen species, and induction of defense-related genes. The molecular role of RanBP1 in plants is discussed in comparison with RanBP1 function in yeast and mammals.

• Journal of Life Science
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• v.13 no.4
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• pp.383-389
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• 2003

A cDNA (SeMIPS) encoding myo-inositol 1-phosphate synthase has been isolated from developing sesame (Sesamum indicum L. cv. Dan-Baek) seeds and its structure and function analyzed. The SeMIPS protein was highly homologous with those from plant species (88-94%), while a much lower degree of sequence homology (60%) was found with that of human. The functional domains commonly found in MIPS protein were identified and their amino acid residues were compared with each other. Northern blot indicated that the expression of the SeMIPS gene might be organ-specifically regulated. A complementation assay based on a yeast mutant system confirmed that the SeMIPS gene encodes a myo-inositol 1-phosphate synthase (MIPS) of sesame by showing functional expression of the SeMIPS cDNA in the yeast mutants containing the disrupted INO1 gene.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.37-44
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• 1991

- I aimed to isolate trans-acting factors involved in the basal expression level of eukaryotic genes. One of the yeast histidine biosynthetic gene, HIS5 was taken as a model for this study. HIS5 gene has a substantial basal level in amino acid rich medium and is derepressed if starved for any single amino acid. The derepression is mediated by cis-acting DNA sequences 5'-TGACTC-3' found in 5' non-transcribed region of the gene and trans-acting factors including GCN4 as positive factor and its negative factor GCDI 7, and GCNZ as a negative factor of GCD17. I first investigated the role of these trans-acting factors in HIS5 basal expression level by using HIS5-pH05 fusion in which expression of pH05 gene encoding inorganic phosphate-repressible acid phosphatase (APase) is regulated by HIS5 promoter. Strain with gcn2 or gcn4 mutation showed 3 to 4 fold lower APase activity than wild type. The level of APase activity was similar in gcn2 and gcn4 mutants. Trans-acting factors involved in basal level were identified by isolating 14 mutants showing increased expression of HISSPH05 fusion from gcn4 background. All the mutants carry a single nuclear recessive mutation and fall into four complementation groups, designated as bell (basal expression level), be12, be23 and be14.

• BMB Reports
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• v.40 no.3
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• pp.349-357
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• 2007

The ubiquitous family of 14-3-3 proteins functions as regulators in a variety of physiological processes. Eight rice 14-3-3 genes, designated OsGF14a through h, were identified from an exhaustive search of the genome database. Comparisons of deduced amino acid sequences reveal a high degree of identity among members of the OsGF14 family and reported Arabidopsis 14-3-3 proteins. A phylogenetic study indicates that OsGF14s contain both $\varepsilon$ and non-$\varepsilon$ forms, which is also confirmed by a structural analysis of OsGF14 genes. Furthermore, transcripts of OsGF14b, OsGF14c, OsGF14d, OsGF14e, OsGF14f and OsGF14g were detected in rice tissues. Their different expression patterns, the different effects of environmental stresses and plant hormones on their transcription levels, and the different complementary phenotypes in yeast 14-3-3 mutants not only indicates that OsGF14s are responsive to various stress conditions and regulated by multiple signaling pathways, but also suggests that functional similarity and diversity coexist among the members of OsGF14 family.

• Journal of Applied Biological Chemistry
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• v.51 no.3
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• pp.102-110
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• 2008

Complementation assay of the urea uptake-defective yeast mutants led to the identification of the Arabidopsis AtTIP4;1 gene encoding the aquaporin. However, its physiological functions still remain elusive. In the present study, histochemical and genetic analyses were performed to understand the physiological roles of AtTIP4;1 in urea uptake. The AtTIP4;1 product was detectible in the roots, but not in the leaves, the stem, and the flower. Its promoter allowed the expression of the $\beta$-glucuronidase reporter gene in the roots and the apical meristem in Arabidopsis. The AtTIP4;1 products were induced under nitrogen-deficient conditions. To investigate the role of the tonoplast intrinsic protein in urea transport and developments, Arabidopsis with the loss- and the gain-of-function mutations by T-DNA insertion in AtTIP4;1 and 35S promoter-mediated overexpression of AtTIP4;1 were identified, respectively. The transfer DNA insertion and the AtTIP4;1-overexpressed plants showed normal growth and development under normal or abiotic stress growth conditions. The urea-uptake studies using $^{14}C$-labeled urea revealed higher accumulation of urea in the AtTIP4;1-overexpressed plants. These results provide evidence that overexpression of AtTIP4;1 leads to the increase in the urea-uptake rate in plants without detectable defects to the growth and development.