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Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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The Individual Discrimination Location Tracking Technology for Multimodal Interaction at the Exhibition (전시 공간에서 다중 인터랙션을 위한 개인식별 위치 측위 기술 연구)

  • Jung, Hyun-Chul;Kim, Nam-Jin;Choi, Lee-Kwon
    • Journal of Intelligence and Information Systems
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    • v.18 no.2
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    • pp.19-28
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    • 2012
  • After the internet era, we are moving to the ubiquitous society. Nowadays the people are interested in the multimodal interaction technology, which enables audience to naturally interact with the computing environment at the exhibitions such as gallery, museum, and park. Also, there are other attempts to provide additional service based on the location information of the audience, or to improve and deploy interaction between subjects and audience by analyzing the using pattern of the people. In order to provide multimodal interaction service to the audience at the exhibition, it is important to distinguish the individuals and trace their location and route. For the location tracking on the outside, GPS is widely used nowadays. GPS is able to get the real time location of the subjects moving fast, so this is one of the important technologies in the field requiring location tracking service. However, as GPS uses the location tracking method using satellites, the service cannot be used on the inside, because it cannot catch the satellite signal. For this reason, the studies about inside location tracking are going on using very short range communication service such as ZigBee, UWB, RFID, as well as using mobile communication network and wireless lan service. However these technologies have shortcomings in that the audience needs to use additional sensor device and it becomes difficult and expensive as the density of the target area gets higher. In addition, the usual exhibition environment has many obstacles for the network, which makes the performance of the system to fall. Above all these things, the biggest problem is that the interaction method using the devices based on the old technologies cannot provide natural service to the users. Plus the system uses sensor recognition method, so multiple users should equip the devices. Therefore, there is the limitation in the number of the users that can use the system simultaneously. In order to make up for these shortcomings, in this study we suggest a technology that gets the exact location information of the users through the location mapping technology using Wi-Fi and 3d camera of the smartphones. We applied the signal amplitude of access point using wireless lan, to develop inside location tracking system with lower price. AP is cheaper than other devices used in other tracking techniques, and by installing the software to the user's mobile device it can be directly used as the tracking system device. We used the Microsoft Kinect sensor for the 3D Camera. Kinect is equippedwith the function discriminating the depth and human information inside the shooting area. Therefore it is appropriate to extract user's body, vector, and acceleration information with low price. We confirm the location of the audience using the cell ID obtained from the Wi-Fi signal. By using smartphones as the basic device for the location service, we solve the problems of additional tagging device and provide environment that multiple users can get the interaction service simultaneously. 3d cameras located at each cell areas get the exact location and status information of the users. The 3d cameras are connected to the Camera Client, calculate the mapping information aligned to each cells, get the exact information of the users, and get the status and pattern information of the audience. The location mapping technique of Camera Client decreases the error rate that occurs on the inside location service, increases accuracy of individual discrimination in the area through the individual discrimination based on body information, and establishes the foundation of the multimodal interaction technology at the exhibition. Calculated data and information enables the users to get the appropriate interaction service through the main server.

Estimation of Mean Surface Current and Current Variability in the East Sea using Surface Drifter Data from 1991 to 2017 (1991년부터 2017년까지 표층 뜰개 자료를 이용하여 계산한 동해의 평균 표층 해류와 해류 변동성)

  • PARK, JU-EUN;KIM, SOO-YUN;CHOI, BYOUNG-JU;BYUN, DO-SEONG
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.24 no.2
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    • pp.208-225
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    • 2019
  • To understand the mean surface circulation and surface currents in the East Sea, trajectories of surface drifters passed through the East Sea from 1991 to 2017 were analyzed. By analyzing the surface drifter trajectory data, the main paths of surface ocean currents were grouped and the variation in each main current path was investigated. The East Korea Warm Current (EKWC) heading northward separates from the coast at $36{\sim}38^{\circ}N$ and flows to the northeast until $131^{\circ}E$. In the middle (from $131^{\circ}E$ to $137^{\circ}E$) of the East Sea, the average latitude of the currents flowing eastward ranges from 36 to $40^{\circ}N$ and the currents meander with large amplitude. When the average latitude of the surface drifter paths was in the north (south) of $37.5^{\circ}N$, the meandering amplitude was about 50 (100) km. The most frequent route of surface drifters in the middle of the East Sea was the path along $37.5-38.5^{\circ}N$. The surface drifters, which were deployed off the coast of Vladivostok in the north of the East Sea, moved to the southwest along the coast and were separated from the coast to flow southeastward along the cyclonic circulation around the Japan Basin. And, then, the drifters moved to the east along $39-40^{\circ}N$. The mean surface current vector and mean speed were calculated in each lattice with $0.25^{\circ}$ grid spacing using the velocity data of surface drifters which passed through each lattice. The current variance ellipses were calculated with $0.5^{\circ}$ grid spacing. Because the path of the EKWC changes every year in the western part of the Ulleung Basin and the current paths in the Yamato Basin keep changing with many eddies, the current variance ellipses are relatively large in these region. We present a schematic map of the East Sea surface current based on the surface drifter data. The significance of this study is that the surface ocean circulation of the East Sea, which has been mainly studied by numerical model simulations and the sea surface height data obtained from satellite altimeters, was analyzed based on in-situ Lagrangian observational current data.