This study re-examined the issues of Korean binding domain investigated in the previous studies (Kim and Yoon 2009, Kim 2013) by testing the validity of Tensed S Condition (TSC) in Korean binding. Hypothesizing that Korean TSC-violating anaphors are indeed exempt anaphors, the current study is designed to fix the problems of the previous studies. Twenty seven Korean native speakers were tested over Acceptability Judgment Task combined with Interpretation Task composed of 155 Korean sentences representing various binding conditions and Korean local anaphors. Overall results showed that Korean native speakers treat sentences with TSC-violating anaphors similarly to those with SSC-violating anaphors rather than the sentences representing local binding, which means that TSC-violating local anaphors are licensed as exempt anaphors in Korean, not core anaphors.

Spatial- and temporal-specific expression patterns are primarily regulated at the transcriptional level by the promoter. Therefore, it is important to determine the binding motifs of transcription factors to understand the networks associated with embryogenesis. Here, we used a protein-binding microarray (PBM) to determine the binding motif of OsSMF1, which is a basic leucine zipper transcription factor that is involved in the regulation of rice seed maturation. OsSMF1 (previously called RISBZ1) is known to interact with GCN4 motifs (TGA(G/C)TCA) to regulate seed storage proteins (SSPs). In addition, OsSMF1 (also known as OsbZIP58) functions as a key regulator of starch synthesis in the rice seed. Quadruple 9-mer-based PBM (Q9-PBM) and electrophoretic mobility shift assay (EMSA) experiments revealed that OsSMF1 binds to the ACGT (CCACGT(C/G)), GCN4 (TGA(G/C)TCA), and GCN4-like (GGATGAC) motifs with Kd values of 0.3353 μM, 0.6458 μM, and 1.117 μM, respectively. We also identified 60 putative OsSMF1 target genes using a combination of data from expression microarrays and RiceArrayNet (RAN) analysis. Of these OsSMF1 target genes, 20, 22, and 17 genes contained ACGT, GCN4, and GCN4-like motifs within the 2-kb promoter region, respectively. In addition to known target genes, we also identified 35 potential OsSMF1 target genes that have not been previously described in immature seeds. We also confirmed that OsSMF1 directly regulates Os03g0168500 (thioredoxin-related protein), RPBF, NAC6, and two hypothetical proteins (Os12g0621600 and Os11g0582400) in vivo. This study suggests that OsSMF1 functions in a wide range of seed development processes with specific binding affinities for three DNA binding motifs

내염성과 관련 있는 벼의 CBF4 유전자(OsCBF4)를 벼에서 과발현 시켜 염 스트레스 조건에서 저항성이 증진된 내염성 유전자원을 개발하였다. RT-PCR을 수행하여 분리한 OsCBF4 유전자는 1,429 bp 크기의 뉴클레오티드로 274개의 아미노산으로 구성되었고, 벼의 다른 CBF 유전자와 33~49% 상동성을 나타내었다. 형질전환 식물체는 PCR과 Southern분석으로 OsCBF4 유전자의 벼 게놈 내 도입을 확인하였다. 전이 유전자는 벼 게놈 내에 1~4사본이 도입되었고, 선발된 형질전환 계통 모두에서 전이 유전자가 강하게 발현되었다. 염 스트레스 조건에서 형질전환 식물체는 비 형질전환 벼 보다 생육 정도가 양호하였으며, 특히 CBF4-10 계통은 염 처리 후 많은 식물체가 살아남았다. Real-time PCR 분석 결과 CBF4-10 계통은 120 mM NaCl 처리 시 전이 유전자 OsCBF4 전사체 발현이 잎에서 무처리 대비 약 3배 이상 증가하였다. 결론적으로 일반 벼에 도입된 OsCBF4 전이 유전자는 내염성 증진 기능이 있으며, OsCBF4 전이 유전자의 발현이 높은 형질전환 벼 계통은 내염성 벼 육종 소재로 이용할 수 있을 것으로 평가된다.

Binding Conditions A and B are quite useful generalizations, but they raise many fundamental questions such as 'why must an anaphor be bound within a local domain, but not outside of it?', and 'why does a pronominal obey an almost opposite constraint?' and 'how is the local domain defined?'. This article explores the possibility of providing answers to those fundamental questions by assuming that binding conditions have both lexical and syntactic aspects. I claim that self is a reflexive predicate that requires its two arguments to be co-indexed, whereas a non-reflexive predicate like like requires its two arguments not to be co-indexed.

In order to utilize waste wood chip for pavement, a polyurethane resin that is both eco-friendly and suitable for bindingwood chip was developed as the binder, and workability was examined through laboratory experiment for characteristicsof waste wood chip mixture using the polyurethane resin and through test pavement on the field. The new resin was aVOC reduction type free from plasticizer and solvent classified as endocrine-disrupting chemicals and environmentalhazardous substances, and NCO equivalents were set at 8, 9, 10 and 11% by modifying the polyisocyanate-polyol ratio.Laboratory experiment showed that polyurethane resin with NCO equivalent of 9% and 10% had excellent characteristicsas binder for waste wood chip. In the field experiment applying waste wood chip and polyurethane resin in the massratio of 1:0.8, tensile strength of the pavement system was about 30% higher than that using polyurethane resin currentlybeing sold, and permeability coefficient and elasticity thereof were the same as that using the resin currently being sold.Also, examination of compaction methods for waste wood chip pavement system showed that non-heating hand rollerand compactor had the problem of the “waste wood chip - resin mixture” sticking to the roller during the compaction but that heating hand roller had excellent workability and could achieve good planation surface relatively easily.

시멘트 내 6가 크롬은 피부질환이나 암을 유발할 수 있는 유해한 인체에 유해한 이온으로 잘 알려져 있다. 본 연구에서는 사람에게 영향을 주는 시멘트 내 6가 크롬의 정량적 분석을 하고자 시멘트 및 시멘트 경화체 내 6가 크롬의 고정화에 대해 평가하였다. 국내에서 생산되는 일반 포틀랜드 시멘트 3종과 플라이애쉬, 고로슬래그 미분말, 실리카퓸을 사용하여 수용성 및 산가용성 6가 크롬을 분광광도법으로 측정하였다. 측정결과, 시멘트 내 수용성 6가 크롬의 농도는 10.5-18.9 mg/kg-cement 범위였으며 혼화재료 내 수용성 6가 크롬의 양은 매우 적게 측정되었다. 시멘트 내 산가용성 6가 크롬의 농도는 172.4-318.2 mg/kg-cement 범위로 측정되었으며 수용성 6가 크롬에 비해 증가하였다. 그러나 크롬의 pH에 의존적인 용해 특성에 따라 용매의 pH가 저감된다고 하여 산가용성 6가 크롬의 농도가 항상 크게 측정되지는 않았다. 시멘트 수화 후 수용성 6가 크롬은 2.0 mg/kg-cement 정도로 감소하였으며 이는 크롬산-에트링게이트 생성에 의한 결과임을 확인할 수 있었다.

The present study concerns the properties of cement-free concrete using binding capacity of cement paste. The cement-free was casted with alkali-activators(KOH, NaOH, and CaOH2) by weight of binder. The properties of cement-free concrete was studied compare to that of OPC. It was found that an increase in the chloride concentration resulted in a decrease the binding capacity of OPC.

As byproducts of chicken slaughtering, chicken feathers are produced and mostly discarded without proper treatment, which results in serious environment pollution. Therefore, the appropriate treatment and utilization of chicken feathers are needed. In particular, chicken feathers can be used as protein sources for the preparation of protein hydrolysates, considering that chicken feathers have a large amount of proteins. In this study, chicken feather protein hydrolysates were prepared and their iron-binding peptides were isolated. Chicken feather protein was extracted from feathers of slaughtered chicken, and its hydrolysates were prepared via hydrolysis with Flavourzyme for 8 h. Then the chicken feather protein hydrolysates were ultra-filtered to obtain small peptide fractions and fractionated using Q-Sepharose and Sephadex G-15 columns to isolate their iron-binding peptides. Two major fractions were produced from each of the Q-Sepharose ion exchange chromatography and the Sephadex G-15 gel filtration hromatography. Among the fractions, the peptide fraction with a high iron-binding activity level, F12, was isolated. These results suggest that chicken feather protein hydrolysates can be used as iron supplements.

Lhx8 (LIM homeobox 8) gene encodes a LIM homeodomain transcriptional regulator that is preferentially expressed in germ cells and critical for mammalian folliculogenesis. However, Lhx8 DNA binding sequences are not characterized yet. We aimed to identify and characterize a cis-acting sequence of germ-cell specific transcriptional factor, Lhx8. To identify Lhx8 DNA binding element, Cyclic Amplification of Sequence Target (CAST) Analysis was performed. Electrophoretic Mobility Shift Assay (EMSA) was processed for the binding specificity of Lhx8. Luciferase assay was for the transcriptional activity of Lhx8 through identified DNA binding site. We identified a putative cis-acting sequence, TGATTG as Lhx8 DNA binding element (LBE). In addition, Lhx8 binds to the LBE with high affinity and augments transcriptional activity of luciferase reporter driven by artificial promoter containing the Lhx8 binding element. These findings indicate that Lhx8 directly regulates the transcription of genes containing Lhx8 binding element in oocytes during early folliculogenesis.

Glycan epitopes of cellular glycoconjugates act as versatile biochemical signals, and this sugar coding plays an important role in cell-to-cell recognition processes. In the present study, our aims were to determine the distribution of sperm receptors with activity for fucosyl- and galactosyl glycans and to address whether mono sugar neoglycoproteins functionally mimic the binding between zona pellucida (ZP) glycoproteins and sperm. In mouse epididymal spermatozoa with intact acrosomes, fucopyranosyl bovine serum albumin (BSA-Fuc) bound to the segment of the acrosome, the equatorial segment, and the postacrosome region of the sperm head. Galactosyl BSA (BSA-Gal) binding activity was similar to that of BSA-Fuc, but was weaker. In acrosome-reacted spermatozoa treated with the Ca2+ ionophore A23187, BSA-Fuc binding was lost in the apical segment of the acrosome but remained in the equatorial segment and postacrosome regions. BSA-Gal binding to the equatorial region was increased. In the presence of 2.5 μg/ml BSA-Fuc, in vitro sperm - ZP binding was significantly decreased, indicating that fucosyl-BSA functionally mimics ZP glycoproteins during sperm-egg ZP interactions. At the same concentration, BSA-Gal was not effective. Fucosyl BSA which efficiently inhibited the sperm-ZP binding can mimic the ZP glycoconjugate and has potential for use as a sperm fertility control agent in mouse.

본 연구에서는 황토에 시멘트나 유기계 접착제를 첨가하지 않은 천연재료 황토결합재를 사용하여 황토경화체를 제작하고, 배합조건에 따른 강도성상을 평가하였다. 황토와 석회의 결합재에 천연재료를 첨가할 경우 사용한 천연재료는 모두 물리적 성능을 개선하는 효과가 있었다. 천연재료 중에서 석회는 황토경화체의 물성을 증가시키는데 가장 큰 영향을 미치고 있다. 황토경화체의 물리적 특성은 적용한 배합비 중에서 W/B 45%, 단위수량 285kg/m3, 석회첨가율 60%일 때 가장 우수하게 나타나고 있다.

The surveying of binding affinity between a particular transcription factor and DNA motifs is important in order to understand the developmental specific gene expression and regulatory networks of an organism. The microarray-based technologies (protein-binding microarrays; PBMs) provide useful predictions for understanding the transcriptional regulatory code in a genome-wide manner. The PBM was designed in such a way that target probes were synthesized as quadruples of all possible 9-mer combinations, named Q9-UPBM. Also, we developed rice promoter PBM (RPBM) using 19,480 rice promoter sequences containing 40 bp long probe with overlapping 20 bp (cover 1kb from 5’ upstream). We applied RISBZ1 protein, an endosperm specific basic leucine zipper transcription factor, to compare binding site specificities between Q9-UPBM and RPBM and find directly regulated promoter regions through the RPBM. Several cis-elements; Prolamin box (TGTAAAG), GCN4 motif (TGA(G/C)TCA), AACA motif (AACAAAA), and ACGT motif, are highly conserved in the promoters of cereal seed storage protein genes, and play a central role in controlling endosperm specific expression during seed maturation. Characterization of cis-elements and TFs has been performed on many storage protein genes of several crop plants, but the mechanisms are still poorly understood. Two chips provide RISBZ1 could bind to ACGT motif such as a CCACGTCA site and GGATGAC site as well as GCN4 motif known binding site. In RPBM binding affinity to CCACGTCA was highly significant, compared to GGATGAC site. The difference might be caused by the biased presence of specific promoter rather than Q9-UPBM. Also our results will provide direct insight into the importance of combinatorial interplay between cis-elements in regulating the expression of seed storage protein genes.

MYB-like domain (MLD) gene is a transcription factor that plays a diverse role in plant development and in response to abiotic stresses. In this study, we isolated and developed CaMV35S::OsMLD rice lines and determined its expression pattern under abiotic stresses. It has Myb_CC_LHEQLE superfamily similar to most transcription factor genes but with a very unique binding domain of SHLQKYR in the C-terminal region. Overexpressing rice lines showed enhanced tolerance to salinity with elevated mRNA transcript. Additionally, mRNA transcripts were up-regulated by ABA, H2O2 and dehydration stresses. Further investigation in the enhanced tolerance to salinity showed an increased accumulation of proline and a decreased in malondialdehyde contents indicating that OsMLD gene may be involved in the regulation of proline and osmolytes during abiotic stresses. These results showed that OsMLD gene could be used in the development of rice intended for soil with salinity-related problem.

Cold stress at the seedling stage is a major threat to rice production. Cold tolerance is controlled by complex genetic factors. We used an F7 recombinant inbred line (RIL) population of 123 individuals derived from the cross of a cold-tolerant japonica and a cold-sensitive indica cultivars, for QTL mapping. Phenotypic evaluation of the parents and RILs in an 18/8oC (day/night) cold-stress regime showed continuous variations for cold tolerance or sensitivity. Six QTLs for seedling cold tolerance were identified on chromosomes 1, 2, 4, 10, and 11 with percent phenotypic variation (R2) ranging from 6.1% to 16.5%. Three main-effect QTLs (qSCT1, qSCT4, and qSCT11) were detected in all cold-tolerant RILs which explained high sum of phenotypic variation (SPV) ranging from 27.1% to 50.6%. Two QTLs (qSCT1 and qSCT11) on chromosomes 1 and 11 were fine mapped. The marker In1-c3 from ORF LOC_Os01g69910 of the BAC clone B1455F06 encoding calmodulin-binding transcription activator (CAMTA) and another marker, In11-d1 from ORF LOC_Os11g37720 (Duf6 gene) of the BAC clone OSJNBa0029K08, co-segregated with seedling cold tolerance. These two InDel markers amplified 241-bp and 158-bp alleles, respectively, in cold-tolerant RILs, and in the cold-tolerant donor Jinbu, which were absent in cold-sensitive parent BR29 and cold-sensitive RILs.

eIF4E family is well known for recessive resistance gene of potyvirus in many crops. And Turnip mosaic virus (TuMV) is one of the major viruses in Brassicaceae crops which belong to the genus Potyvirus. To elucidate the key amino acids in the interaction between TuMV VPg and Brassica eIF(iso)4E, amino acids of eIF(iso)4E were mutated. Seven amino acids in cap binding pocket were chose for the candidate amino acid that may play a role in the interaction of TuMV VPg. We demonstrated that a single amino acid mutation in cap binding pocket of Brassica eIF(iso)4E can abolish the interaction with TuMV VPg. eIF(iso)4E which has a mutation at each W49, W95 and K150 positions impaired in its interaction with VPg prominently according to the yeast two hybrid analysis. Complementation of an eIF4E knockout yeast strain by mutated eIF(iso)4E proteins showed that all eIF(iso)4E mutants were able to complement eIF4E of yeast. To find out if these mutations affect the susceptibility of Chinese cabbage, transformant analysis was performed. eIF(iso)4E W95L, W95L/K150E and susceptible wild type were over-expressed in susceptible Chinese cabbage. According to the TuMV screening result of T1 and T2 transformants, over-expression of the eIF(iso)4E mutants showed resistance to four TuMV strains (CHN2, 3, 4 and 5). Our results support that the mutations in eIF(iso)4E may control the broad spectrum TuMV resistance.

To characterize CBF/DREB1-homologue in rice, nine OsDREB1 genes have been identified and characterized in this lab. Among these, it was shown that OsDREB1D was induced by drought and slightly by cold stress. We found that OsDREB1A, -1D, and -1E could up-regulate OsDhn1:LUC construct in transactivation assay using rice protoplasts. Transgenic rice plants overexpressing OsDREB1D under the maize ubiquitin promoter (Ubi:OsDREB1D) revealed an enhanced stress tolerance to drought. We also generated transgenic rice of OsDREB1D under OsPOX1 promoter (OsPOX1:OsDREB1D), which is cold stress inducible preferentially in the reproductive organs of rice. We are currently examining the mechanism of the enhanced tolerance of the transgenic plants to drought stress using both molecular physiological and biochemical techniques.

선행연구에서 본 연구자는 IEX-1 단백질이 난소 암세포에서 세포사멸(apoptosis)을 유도하는 기능을 수행함을 확인하였으나, 세포사멸과 세포생존의 여러 단계에서 어떠한 신호전달 체계로 IEX-1이 작용하는지는 정확히 알지 못하고 있다. 따라서 IEX-1 단백질과 결합하는 새로운 단백질을 찾기 위해 yeast two-hybrid system을 이용하였다. 그 결과 IEX-1이 여러 다양한 인간 암 세포에서 세포사멸을 유도하는 CATHEPSIN B 단

MYB-like domain (MLD) gene is a transcription factor that plays a diverse role in plant development and in response to abiotic stresses. In this study, we isolated and developed CaMV35S::OsMLD rice lines and determined its expression pattern under abiotic stresses. The MLD has Myb_CC_LHEQLE superfamily similar to most transcription factor genes but with a very unique binding domain of SHLQKYR in the C-terminal region. Overexpressing rice lines showed enhanced tolerance to salinity with elevated mRNA transcript. Additionally, mRNA transcripts were up-regulated by ABA, H2O2 and dehydration stresses. Further investigation in the enhanced tolerance to salinity showed an increased accumulation of proline and a decreased in malondialdehyde contents indicating that OsMLD gene may be involved in the regulation of proline and osmolytes during abiotic stresses. These results showed that OsMLD gene could be used in the development of rice intended for soil with salinity-related problem.