• Proceedings of the Botanical Society of Korea Conference
• /
• 1996.07a
• /
• pp.61-74
• /
• 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.

• Journal of Microbiology
• /
• v.45 no.2
• /
• pp.158-167
• /
• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Journal of the Institute of Electronics Engineers of Korea SD
• /
• v.45 no.7
• /
• pp.61-68
• /
• 2008

In this paper, we propose an efficient hardware architecture and algorithm to increase an encoding process rate and implement a hardware for CABAC (Context Adaptive Binary Arithmetic Coding) which is used with one of the entropy coding ways for the latest video compression technique, H.264/AVC (Advanced Video Coding). CABAC typically provides a better high compression performance maximum 15% compared with CAVLC. However, the complexity of operation of CABAC is significantly higher than the CAVLC. Because of complicated data dependency during the encoding process, the complexity of operation is higher. Therefore, various architectures were proposed to reduce an amount of operation. However, they have still latency on account of complicated data dependency. The proposed architecture has two techniques to implement efficient pipeline architecture. The one is quick calculation of 7, 8th bits used to calculate a probability is the first step in Binary arithmetic coding. The other is one step reduced pipeline arcbitecture when the type of the encoded symbols is MPS. By adopting these two techniques, the required processing time was reduced about 27-29% compared with previous architectures. It is designed in a hardware description language and total logic gate count is 19K using 0.18um standard cell library.

• Journal of Life Science
• /
• v.13 no.4
• /
• pp.529-534
• /
• 2003

This study was done to investigated the phytosterol compositions of safflower (Carthamus tinctorius L.) seed. The phytoestrogen activity was also determined using CAT-ELISA Kit in ethanol extract of safflower seed. The phytosterol of safflower seeds was identified using gas chromatography-mass spectrometry after saponification of the oils. The phytosterol content and composition of safflower seed oils were 4% and identified stigmast-5-en-3-ol (3$\beta$, 24S)-form, ${\gamma}$-sitosterol (clionasterol) with Wiley MS spectrum library. The synergistic effect of human estrogen receptor (hER) has been investigated using a minimal chimeric promoters composed of the TATA region of the adenovirus-2 major late promoter (A22MLP) and two consensus perfectly polindromic Xenopus vitellogenin A2 gene estrogen responsive elements (XVEREl19). Transient transfection experiments in tile human breast adenocarcinoma cell line MCF-7, which is known to express the estrogen receptor endogenously, revealed that phytoestrogen from Carthamus tinctorius L. acts as estrogen. We have observed the transcriptional activities stimulated methanol and ethanol extract of safflower seed in MCF-7, were 0.43 and 0.37 respectively, compared to that by $\beta$-estradiol as 1.0. Our data showed that safflower seeds have estrogenic activity methanol and ethanol extracts and ethanol lower than that of $\beta$-estradiol. This result provides the first evidence that the beneficial effect of safflower seeds may be mediated, at least in part, by the stimulating effect of phytoestrogen ell bone-protecting.

• Radiation Oncology Journal
• /
• v.17 no.2
• /
• pp.151-157
• /
• 1999

Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

• Animal cells and systems
• /
• v.1 no.3
• /
• pp.521-526
• /
• 1997

We isolated a mouse nck cDNA from the thymus cDNA expression library. The cDNA encodes a 377 amino acid protein and displays 97% amino acid sequence identity to human oncogenic protein nck, which is composed almost exclusivelv of three src homology 3 (SH3) domains and one SH2 domain. The sequence analysis also showed that the isolated cDNA is the mouse counterpart of the human nck and different from the mouse grb4, which has been reported to be highly similar to the human nck and, therefore considered as a mouse nck, Northern blot analysis showed that the transcript of the gene was 1.8 kb and was highly expressed in the testis, thymus, and brain but moderately in the liver and lymph node. Western blot analysis showed that the size of the protein was about 47 kDa. Overexpression of the mouse Nck transformed a mouse fibroblast cell line, NIH3T3. The results clearly indicate that normal nck gene has transforming ability and provide an argument against a suggested possibility that the transforming ability of the human nck gene is due to a mutation(s) in the gene.

• The Journal of Korean Institute of Communications and Information Sciences
• /
• v.30 no.4C
• /
• pp.283-289
• /
• 2005

In this paper, we propose two efficient symbol detection algorithms for space-frequency OFDM (SF-OFDM) transmit diversity scheme. When the number of sub-carriers in SF-OFBM scheme is small, the interference between adjacent sub-carriers may be generated. The proposed algorithms eliminate this interference in a parallel or sequential manlier and achieve a considerable performance improvement over the conventional detection algorithm. The bit error rate (BER) performance of the proposed detection algorithms is evaluated by the simulation. In the case of 2 transmit and 2 receive antennas, at $BER=10^{-4}$ the proposed algorithms achieve the gain improvement of about 3 dB. The symbol detectors with the proposed algorithms are designed in a hardware description language and synthesized to gate-level circuits with the $0.18{\mu}m$ 1.8V CMOS standard cell library. With the division-free architecture, the proposed SF-OFDM-PIC and SF-OFDM-SIC symbol detectors can be implemented using 140k and 129k logic gates, respectively.

• Journal of the Korea Institute of Information and Communication Engineering
• /
• v.14 no.3
• /
• pp.715-722
• /
• 2010

This paper describes a hardware design of hash function processor which implements Korean Hash Algorithm Standard HAS-160. The HAS-160 processor compresses a message with arbitrary lengths into a hash code with a fixed length of 160-bit. To achieve high-speed operation with small-area, arithmetic operation for step-operation is implemented by using a hybrid structure of 5:3 and 3:2 carry-save adders and carry-select adder. It computes a 160-bit hash code from a message block of 512 bits in 82 clock cycles, and has 312 Mbps throughput at 50 MHz@3.3-V clock frequency. The designed HAS-160 processor is verified by FPGA implementation, and it has 17,600 gates on a layout area of about $1\;mm^2$ using a 0.35-${\mu}m$ CMOS cell library.

• Journal of the Institute of Electronics Engineers of Korea SD
• /
• v.49 no.8
• /
• pp.63-72
• /
• 2012

In this paper, we proposed a parallel architecture of line & block edge filter for high-performance H.264/AVC deblocking filter for Quad Full High Definition(Quad FHD) video real time processing. To improve throughput, we designed 4-parallel block edge filter with 16 line edge filter. To reduce internal buffer size and processing cycle, we scheduled 4-parallel zig-zag scan order as deblocking filtering order. To avoid data conflicts we placed 1 delay cycle between block edge filtering. We implemented interleaving buffer, as internal buffer of block edge filter, to sharing buffer for reducing buffer size. The proposed architecture was simulated in 0.18um standard cell library. The maximum operation frequency is 108MHz. The gate count is 140.16Kgates. The proposed H.264/AVC deblocking filter can support Quad FHD at 113.17 frames per second by running at 90MHz.

• Journal of Advanced Navigation Technology
• /
• v.14 no.3
• /
• pp.328-335
• /
• 2010

In this paper, an efficient algorithm and area-efficient hardware architecture have been proposed for $2{\times}2$ MIMO-OFDM based WLAN baseband modem with two transmit and two receive antennas. To enhance the performance of the receiver, the efficient timing synchronization algorithm and symbol detector based on MML algorithm are presented. Also, by sharing the hardware block with multi-stage pipeline structure and using the complex multiplier based on polar-coordinate, the complexity of the proposed architecture is dramatically decreased. The proposed area-efficient hardware design was designed in hardware description language (HDL) and synthesized to gate-level circuits using 0.13um CMOS standard cell library. As a result, the complexity of the proposed modem receiver is reduced by 56% over the conventional architecture.