• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.174-185
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• 2010

Full-length cDNAs are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. To elucidate the functions of a large population of Chinese cabbage (Brassica rapa) genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the full-length cDNA Over-eXpresser (FOX) gene hunting system. With oligo dT column it purify the each mRNA from the flower organs, leaf and stem tissue. And about 120,000 cDNAs from the library were transformed into $\lambda$-pFLCIII-F vector. Of which 115,000 cDNAs from the library were transformed into T-DNA binary vector, pBigs for transformation study. We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. Full-length Chinese cabbage cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Selfed seeds were harvested from transgenic Arabidopsis. We had selected 2,500 transgenic plants by hygromycin antibiotic tolerant test, and obtained a number of transgenic mutants. Each transgenic Arabidopsis was investigated in morphological changes, fertility and leaf colour. As a result, 285 possible morphological mutants were identified. Introduced cDNA was isolated by PCR amplification of the genomic DNA from the transgenic mutants. Sequencing result and BLAST analysis showed that most of the introduced cDNA were complete cDNAs and functional genes. Also, we examined the effect of Bromelain on enhancing resistance to soft rot in transgenic Chinese cabbage 'Osome'. The bromelain gene identified from FOX hunting system was transformed into Chinese cabbage using Agrobacterium methods. Transformants were screened by PCR, then RT-PCR and real time PCR were performed to analyze gene expression of cysteine protease in the T1 and T2 generations. The anti-bacterial activity of bromelain was tested in Chinese cabbages infected with soft rot bacteria. The results showed that the over-expressed bromelain gene from pineapple conferred enhanced resistance to soft rot in Chinese cabbage.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.452-458
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• 1996

A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

• Journal of Microbiology and Biotechnology
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• v.18 no.3
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• pp.410-416
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• 2008

The gene SfXyn10, which encodes a protease-resistant xylanase, was isolated using colony PCR screening from a genomic library of a feather-degrading bacterial strain Streptomyces fradiae var. k11. The full-length gene consists of 1,437bp and encodes 479 amino acids, which includes 41 residues of a putative signal peptide at its N terminus. The amino acid sequence shares the highest similarity (80%) to the endo-1,4-${\beta}$-xylanase from Streptomyces coelicolor A3, which belongs to the glycoside hydrolase family 10. The gene fragment encoding the mature xylanase was expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to homogeneity by acetone precipitation and anion-exchange chromatography, and subsequently characterized. The optimal pH and temperature for the purified recombinant enzyme were 7.8 and $60^{\circ}C$, respectively. The enzyme showed stability over a pH range of 4.0-10.0. The kinetic values on oat spelt xylan and birchwood xylan substrates were also determined. The enzyme activity was enhanced by $Fe^{2+}$ and strongly inhibited by $Hg^{2+}$ and SDS. The enzyme also showed resistance to neutral and alkaline proteases. Therefore, these characteristics suggest that SfXyn10 could be an important candidate for protease-resistant mechanistic research and has potential applications in the food industry, cotton scouring, and improving animal nutrition.

• Korean Institute of Interior Design Journal
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• v.14 no.6 s.53
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• pp.177-184
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• 2005

The 21st Century Museum of Contemporary Art, Kanazawa as a regional public culture facility is the place where it is possible to think over the roles and meanings of a museum. The urn of this research is to provide information for exploring a desirable direction of the design of a regional museum by analyzing the architectural meanings and public property of the 21st Century Museum of Contemporary Art, Kanazawa. In addition, we arranged the totes and meanings of a regional museum and the concept of the public property in architecture. We also analyzed the architectural quality and the expression of the public property in this museum, as well as the value and meaning of a museum through field researches. The results of this research are as follows. First, this museum breaks with the existing stereotyped Idea to be a space open to the citizens. It secures a wide public space and provides the convenience of access and every facility for citizens. Second, the floor plan is divided into the interchanging area for citizens and an exhibition area; the outskirts of the inside of the museum are planned to be used free of charge for an interchanging space of citizens as a public-owned space of citizens. Third, the public-owned space of citizens includes a citizen exhibition hall, an art library, a kids studio and rest space, which ate all designed lot everyone to enjoy freely. Last, the exterior shape of the museum is simple but harmonize with the surroundings. It has an unique shape showing the local identity, and its most special properties are the convenience of access, introduction of a free space inside, and security of a huge space for children and citizens.

• Molecules and Cells
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• v.27 no.4
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• pp.449-458
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• 2009

The OsAsr1 cDNA clone was isolated from a cDNA library prepared from developing seed coats of rice (Oryza sativa L.). Low-temperature stress increased mRNA levels of OsAsr1 in both vegetative and reproductive organs. In situ analysis showed that OsAsr1 transcript was preferentially accumulated in the leaf mesophyll tissues and parenchyma cells of the palea and lemma. For transgenic rice plants that over-expressed full-length OsAsr1 cDNA in the sense orientation, the Fv/Fm values for photosynthetic efficiency were about 2-fold higher than those of wild type-segregating plants after a 24-h cold treatment. Seedlings exposed to prolonged low temperatures were more tolerant of cold stress, as demonstrated during wilting and regrowth tests. Interestingly, OsAsr1 was highly expressed in transgenic rice plants expressing the C-repeat/dehyhdration responsive element binding factor 1 (CBF1), suggesting the regulation of OsAsr1 by CBF1. Taken together, we suggest that OsAsr1 gene play an important role during temperature stress, and that this gene can be used for generating plants with enhanced cold tolerance.

• Journal of Korean Library and Information Science Society
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• v.9
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• pp.115-144
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• 1982

In ordinary modern wester expression 'the Renaissance' was used to denote the impact made by dead Hellenism civilization in western Christendom, particularly Italian literary and artistic movement at Northern and central Italy in the late medieval period. However, A.J. Toynbee examined the renaissance from the different aspect of view. In his great work "A Study of History" in vol. IX, he succeeded in establishing the theory of historic civilization encounters in space and in time; and in time, civilization of the present and the past or between dead and infant successor contacts on the analogy of parenthood and sonship in the relation of A n.0, pparentation-and-Affiliation. The distinguished his view of 'Renaissance' was illustrated in the sense of encounters between a grown-up civilization and the 'ghost' of its long-dead predecessor. The renaissances (by the process of evocation of ghost of its parent society) has not only one single aspect of literary and artistic field but also in politics, law, science and philosophy, languages and literatures and visual arts, and religion. The main theme of this study is to examine the development of libraries and its historical meaning through Toynbee's literary renaissance. His renaissance of Languages and Literatures has three typical steps: They are: 1st step-to restive the dead literature's remains: 2nd step-to remaster their meaning: 3rd step-to reproduce them in counterfeits... Through its first and second steps, collecting and editing, annotating by compiling an anthology, thesaurus, lexicon or encyclopedia, and in its third step publishing mostly imitation of classics took place. Toynbee depicted the five outstanding eminent representatives of literary renaissance who had a n.0, ppeared on the state of history down to the time of writing. They are: Assurbanipal, Constantine prophyrogenitus, Yung Lo, K'ang Hsi, and Ch'ien Lung and the last four had all been emperors of imperia rediviva. As the result of the examination of these five emperors with three steps of literary renaissance, the common result may be summarized as follows: 1. Those emperors of imperia rediviva interested in intellectual work and study, they also were deeply involved in collecting classics in an ostensible reason. 2. There were strong political intention of collecting materials as an a n.0, ppeasement policy of civilization by transferring scholars energies to an intellectual field. 3. Under the rulers of a resuscitated universal state, the literary renaissance were a product of political plane and that the total size of collection and work were huge. 4. Since there were strong exercise of sovereign power, an active censorship by distortion and elimination was inevitable. 5. There existed newly developed strained atmosphere between grown-up and long-dead parent civilization, whenever the book collection movement had occurred. 6. Over adhesion to the parent civilization caused imitation of classic work and the creative activities were stagnated.stagnated.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.600-605
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• 1989

For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.23-29
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• 2009

We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

• Journal of Life Science
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• v.14 no.2
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• pp.325-330
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• 2004

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (Homologue of RAD3 gene). The over-expressed HRD3 protein was estimated to be a 75 kDa in size which is in good agreement with the estimated by the nucleotide sequence of the cloned gene. Two-dimensional gel electrophoresis showed that a number of other protein spots dramatically disappeared when the HRD3 protein was overexpressed. The overexpressed RAD3 protein showed a toxicity in E. coli host, suggesting that this protein may be involved in the inhibition of protein synthesis and/or degradation of host protein. To determine which part of HRD3 gene contributes to the toxicity in E. coli, various fusion plasmids containing a partial sequence of HRD3 and lac'Z gene were constructed. These results suggest that the C-terminal domain of HRD3 protein may be important for both toxic effect in E. coli and for its role in DNA repair in S. pombe.