Three isozymes of peptidylarginine deiminase in the chicken: Molecular cloning, characterization, and tissue distribution
Shimizu A.; Handa K.; Honda T.; Abe N.; Kojima T.; Takahara H., Peptidylarginine deiminase (PAD; EC 3.5.3.15) is a post-translational modification enzyme that catalyzes the conversion of protein-bound arginine to citrulline (deimination) in a calcium ion dependent manner. Although PADI genes are widely conserved among vertebrates, their function in the chicken is poorly understood. Here, we cloned and sequenced three chicken PADI cDNAs and analyzed the expression of their proteins in various tissues. Immunoblotting analysis showed that chicken PAD1 and PAD3 were present in cells of several central neuron system tissues including the retina; the chicken PAD2 protein was not detected in any tissue. We expressed recombinant chicken PADs in insect cells and characterized their enzymatic properties. The chicken PAD1 and PAD3 recombinant proteins required calcium ions as an essential cofactor for their catalytic activity. The two recombinant proteins showed similar substrate specificities toward synthetic arginine derivatives. By contrast to them, chicken PAD2 did not show any activity. We found that one of the conserved active centers in mammalian PADs had been altered in chicken PAD2; we prepared a reverse mutant but we did not detect an activity. We conclude that chicken PAD1 and PAD3 might play specific roles in the nervous system, but that chicken PAD2 might not be functional under normal physiological conditions. (C) 2013 Elsevier Inc All rights reserved., ELSEVIER SCIENCE INC
Comparative Biochemistry & Physiology, Part B, 2014年, ［査読有り］

Molecular characterization of a novel armadillo repeat-like protein gene differentially induced by high-salt stress and dehydration from the model legume Lotus japonicus
Kojima T.; Kinoshita M.; Yamada T.; Umezaki S.; Iwaizako M.; Saito Y.; Noguchi K.; Takahara H., 筆頭著者, Armadillo (ARM) repeat proteins have tandem repeats of a degenerate sequence motif required for protein-protein interaction and are distributed widely in eukaryotes. In this study, we isolated and characterized a novel ARM repeat-like protein gene from the model legume Lotus japonicus that is differentially induced by abiotic stress. The gene, LjTDF-5, encodes a hypothetical protein of 369 aa with a protein signature "ARM-type fold". Three-dimensional protein structure was predicted to consist of 23 alpha-helices and no beta-sheets by homology modeling. The LjTDF-5-homologous genes were distributed broadly in the plant kingdom and the C-terminal region, around 60 amino acids in length, was highly conserved in all of the homologs examined although any known functional domains or protein signatures in this region were not detected in silico analyses. Subcellular localization assays revealed that the sGFP-fused LjTDF-5 protein localized to the nuclei of onion epidermis cells, despite the protein not containing a typical nuclear localization signal. In quantitative real-time RT-PCR, the expression of LjTDF-5 was highly induced by 100 mM NaCl in the roots and by dehydration in the shoot, but not by abscisic acid (ABA, 10 mu M). These results suggest that the ARM repeat-like protein LjTDF-5 functions in or around the nucleus in response to high-salt stress and dehydration in L. japonicus., SPRINGER
Plant Molecular Biology Reporter, 2013年, ［査読有り］

An intronic enhancer driven by NF-kB contributes to transcriptional regulation of peptidylarginine deiminase type I gene in human keratinocytes
Ying S.; Kojima T.; Kawada A.; Nachat R.; Serre G.; Simon M.; Takahara H., Peptidylarginine deiminases (PADs) catalyze the conversion of protein-bound arginine to citrulline residues. In human epidermis, where filaggrin is the main deiminated protein, three PADs are detected with specific patterns of expression depending on the keratinocyte (KC) differentiation state. Previous characterizations of the PAD-encoding gene promoters have shown that proximal regulation alone is not sufficient to explain this specificity of expression. In this work, we describe an evolutionarily highly conserved nucleotide segment located in the first intron of the PAD1 gene (PADI1). Luciferase reporter assays showed that it enhances the activity of the PADI1 promoter, in a calcium-and orientation-independent manner. Mutation of a putative NF-kappa B cis-element markedly reduced its enhancer activity, which also confirmed its potential regulatory function. Chromatin immunoprecipitation assays evidenced the binding of both p65 and p50 NF-kappa B subunits to the cis-element, and RNA interference inhibition assays confirmed that NF-kappa B contributes to the PADI1 transcriptional control. Furthermore, the intronic enhancer and promoter of PADI1 potentially interact through chromatin looping, as indicated by chromosome conformation capture assays. Our findings provide evidence that an NF-kappa B-mediated signaling pathway is involved in PADI1 regulation in human epidermal KCs., NATURE PUBLISHING GROUP
Journal of Investigative Dermatology, 2010年, ［査読有り］

Molecular characterization of a novel soybean gene encoding a neutral PR-5 protein induced by high-salt stress
Tachi H.; Fukuda-Yamada K.; Kojima T.; Shiraiwa M.; Takahara H., 責任著者, In this study, we characterized a novel soybean gene encoding a neutral PR-5 protein and compared it to two acidic isoforms of soybean PR-5 protein. This gene, designated as Glycine max osmotin-like protein, b isoform (GmOLPb, accession no. AB370233), encoded a putative protein having the greatest similarity to chickpea PR-5b (89% identity). Unlike the two acidic PR-5, GmOLPa and P21, the protein had a C-terminal elongation responsible for possible vacuolar targeting and after maturation showed a calculated molecular mass of 21.9 kDa with pl 6.0. The 3D models, predicted by the homology modeling, contained four alpha-helixes and 16 beta-strands and formed three characteristic domains. The two acidic PR-5 proteins also showed a 3D structure very similar to GmOLPb, although the electrostatic potential on molecular surface of each PR-5 was significantly different. In the study of the gene expression under conditions of high-salt stress, GmOLPb was highly induced in the leaves of the soybean, particularly in the lower part of a leaf The expression started at 2 h after initiation of the stress and was highly induced between 18-72 h. Gene expression of P21e (protein homologous to P21) was transiently induced by high-salt stress, but took place earlier than the gene expressions of GmOLPa and GmOLPb. Such differential expression was observed also under investigation with methyl jasmonate and salicylic acid. These results suggested that each soybean PR-5 might play a distinctive role in the defensive system protecting the soybean plant against high-salt stress, particularly in the leaves of the soybean. (C) 2008 Elsevier Masson SAS. All rights reserved., ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Plant Physiology and Biochemistry, 2009年, ［査読有り］

Molecular characterization of a novel salt-inducible gene for an OSBP (oxysterol-binding protein)-homologue from soybean
Li D.Y.; Inoue H.; Takahashi M.; Kojima T.; Shiraiwa M.; Takahara H., 責任著者, Oxysterol-binding protein (OSBP) and its homologues constitute a protein family in many eukaryotes from yeast to humans, which are involved in cellular lipid metabolism, vesicle transport and signal transduction. In this study, we characterized a novel salt-inducible gene for an OSBP-homologue from soybean (Glycine max [L.] Merr.). The soybean OSBP-homologous gene, denoted as G. max OSBP (GmOSBP), encoded a 789 aa putative protein with two characteristic domains; the pleckstrin homology (PH) domain and the ligand-binding (LB) domain, in the Nand C-terminus, respectively. The GmOSBP-PH domain showed localization into/around the nucleus in a transient subcellular localization assay. The phylogenetic relationship of the GmOSBP-LB domain to those in other OSBP-homologues suggested that GmOSBP might bind a lipid molecule(s) different from the ligand-candidates found for the human/yeast OSBP-homologues. The GmOSBP gene was constitutively transcribed in all of the soybean organs examined - root, stem and trifoliate leaf - at low levels and was highly induced in all these organs by high-salt stress (300 mM NaCl). Interestingly, gene expression of GmOSBP was also markedly induced in the senesced soybean cotyledon, which contains high levels of a variety of cellular lipids utilized for energy for germination and as membrane components. Therefore, we suggest that GmOSBP may be involved in some physiological reactions for stress-response and cotyledon senescence in the soybean. (c) 2007 Elsevier B.V All rights reserved., ELSEVIER SCIENCE BV
Gene, 2008年, ［査読有り］

Crucial roles of MZF1 and Sp1 in the transcriptional regulation of the peptidylarginine deiminase type I gene (PADI1) in human keratinocytes
Dong S.; Ying S.; Kojima T.; Shiraiwa M.; Kawada A.; Mechin M-C.; Adoue V.; Chavanas S.; Serre G.; Simon; M.; Takahara H., Pepticlylarginine deiminases (PADs) catalyze the conversion of protein-bound arginine residues into citrulline residues in a calcium-dependent manner. The PAD1 gene (PADI1) is expressed in a few tissues, including the epidermis, where the protein is detected with a higher level in the more differentiated keratinocytes. Using quantitative reverse transcription-PCR experiments, we show that PADI1 mRNAs are more abundant in keratinocytes cultured with 1.2 than 0.15 mm calcium. We cloned and characterized the promoter region using human keratinocytes transfected with variously deleted fragments of the 5'-upstream region of PADI1 coupled to the luciferase gene. We found that as few as 195bp upstream from the transcription initiation site were sufficient to direct transcription of the reporter gene. Mutations of MZF1- or Sp1-binding sites markedly reduced PADI1 promoter activity. Chromatin immunoprecipitation assays revealed that MZF1 and Sp1/Sp3 bind to this region in vivo. Furthermore, MZF1 or Sp1 small interfering RNAs (siRNAs) effectively diminished PADI1 expression in keratinocytes cultured in both low- and high-calcium-containing medium. In addition, the expression of MZF1 and PAD1 increased in parallel when normal human epidermal keratinocytes underwent differentiation. These data indicate that MZF1 and Sp1/Sp3 binding to the promoter region drive the PADI1 expression., NATURE PUBLISHING GROUP
Journal of Investigative Dermatology, 2008年, ［査読有り］

NF-Y and Sp1/Sp3 are involved in the transcriptional regulation of the peptidylarginine deiminase type III gene (PADI3) in human keratinocytes
Dong S.J.; Kanno T.; Yamaki A.; Kojima T.; Shiraiwa M.; Kawada A.; Mechin M-.C.; Chavanas S.; Serre G.; Simon M.; Takahara H., Human peptidylarginine deiminase type III gene (PADI3) encodes a crucial post-translational modification enzyme that converts protein-bound arginine residues into citrulline residues. Its expression is restricted to a few cell types, including keratinocytes in the granular layer of the epidermis and in the inner root sheath of hair follicles. In these cells, the enzyme is involved in terminal processing of intermediate filament-binding proteins such as filaggrin and trichohyalin. To study the molecular mechanisms that control the expression of PADI3 in human keratinocytes at the transcriptional level, we characterized its promoter region using human keratinocytes transfected with variously deleted fragments of the 5'-upstream region of PADI3 coupled to the luciferase gene. We found that as few as 129 bp upstream from the transcription initiation site were sufficient to direct transcription of the reporter gene. Electrophoretic mobility-shift and chromatin immunoprecipitation assays revealed that NF-Y (nuclear factor Y) and Sp1/Sp3 (specificity protein 1/3) bind to this region in vitro and in vivo. Moreover, mutation of the Sp1- or NF-Y-binding motif markedly reduced PADI3 promoter activity. Furthermore, Sp1 or NF-YA (NF-Y subunit) small interfering RNAs effectively diminished PADI3 expression in keratinocytes cultured in both low- and high-calcium medium. These data indicate that PADI3 expression is driven by Sp1/Sp3 and NF-Y binding to the promoter region., PORTLAND PRESS LTD
Biochemical Journal, 2006年, ［査読有り］

Molecular cloning and characterization of a novel salt-inducible gene encoding an acidic isoform of PR-5 protein in soybean (Glycine max [L.] Merr.)
Onishi M.; Tachi H.; Kojima T.; Shiraiwa M.; Takahara H., 責任著者, We identified a novel salt-inducible soybean gene encoding an acidic-isoform of pathogenesis-related protein group 5 (PR-5 protein). The soybean PR-5-homologous gene, designated as Glycine max osmotin-like protein, acidic isoform (GmOLPa); accession no, AB116251), encodes a putative polypeptide having an N-terminal signal peptide. The mature GmOLPa protein without the signal peptide has a calculated molecular mass of 21.5 kDa and a pI value of 4.4, and was distinguishable from a known PR-5-homologous gene of soybean (namely P21 protein) through examination of the structural features. A comparison with two intracellular salt-inducible PR-5 proteins, tobacco osmotin and tomato NP24, revealed that GmOLPa did not have a C-terminal extension sequence functioning as a vacuole-targeting motif The GmOLPa gene was transcribed constitutively in the soybean root and was induced almost exclusively in the root during 24 It of high-salt stress (300 mM NaCl). Interestingly, GmOLPa gene expression in the stein and leaf, not observed until 24 h, was markedly induced at 48 and 72 h after commencement of the high-salt stress. Abscisic acid (ABA) and dehydration also induced expression of the GmOLPa gene in the root; additionally, dehydration slightly induced expression in the stem and leaf. In fact, the 5'-upstream sequence of the GmOLPa gene contained several putative cis-elements known to be involved in responsiveness to ABA and dehydration, e.g. ABA-responsive element (ABRE), MYB/MYC, and low temperature-responsive element (LTRE). These results suggested that GmOLPa may function as a protective PR-5 protein in the extracellular space of the soybean root in response to high-salt stress and dehydration. (c) 2006 Elsevier Masson SAS. All rights reserved., ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Plant Physiology and Biochemistry, 2006年, ［査読有り］

Regulation of the expression of peptidylarginine deiminase type II gene (PADI2) in human keratinocytes involves Sp1 and Sp3 transcription factors
Dong S.J.; Kojima T.; Shiraiwa M.; Mechin M-.C.; Chavanas S.; Serre G.; Simon M.; Kawada A.; Takahara H., Peptidylarginine deiminases (PAD) convert protein-bound arginine residues into citrulline residues in a Ca2+ ion-dependent manner. Among the five isoforms (PAD1, 2, 3, 4, and 6) existing in rodents and humans, PAD2 is the most widely expressed in both species, tissues, and organs. In order to study the mechanisms regulating the expression of the human PAD2 gene, PADI2, we characterized its promoter region using transfected human keratinocytes. A series of reporter gene constructions derived from the 2 kb region upstream of the transcription initiation site defined a minimal promoter sequence from nucleotides -132 to -41. This PADI2 region is GC-rich and lacks canonical TATA and CAAT boxes. Investigation of cis-acting elements in the region, further deletion analyses and electrophoretic mobility shift assays using specific antibodies revealed four Sp1-binding sites and identified Sp1 and Sp3 as binding factors important for the promoter activity. These results suggest that Sp1/Sp3 cooperation may provide a mechanism to control the transcription of PADI2., BLACKWELL PUBLISHING INC
Journal of Investigative Dermatology, 2005年, ［査読有り］

Molecular cloning and characterization of a novel soybean gene encoding a leucine-zipper-like protein induced to salt stress
Aoki A.; Kanegami A.; Mihara M.; Kojima T.; Shiraiwa M.; Takahara H., 責任著者, To understand molecular responses to salt stress in soybean (Glycine max [L.] Merr.), we identified 106 salt-inducible soybean genes that expressed differentially at 72 h after 100 mM NaCl treatment using the cDNA-amplified fragment length polymorphism (AFLP) method. The genes were designated as G. max Transcript-Derived Fragments (GmTDFs). Among these genes, we characterized a soybean gene GmTDF-5 that encoded an unknown protein of 367 amino acids. The GmTDF-5 protein was a putative cytosolic protein with two leucine-zipper motifs at the N-terminal and was calculated as 40.7 kDa. Southern blot analysis indicated that GmTDF-5 presents as an intron-less single gene on soybean genome and possibly distributes narrowly throughout the higher plants. By 100 mM NaCl treatment, the gene expression of Gm TDF-5 was induced in the stem and lower-expanded leaf, and the amount of mRNA increased 5.1- and 2.0-fold up to 72 h, respectively. Interestingly, GmTDF-5 expression in the upper-leaf appeared dramatically with 10.0-fold increase at 72 h after the salt stress, but not until 48 h. Hyperosmotic pressure (mannitol treatment) and dehydration also caused the increases similar to NaCl treatment in the levels of GmTDF-5 expression. These results suggest that GmTDF-5 might be a novel cytosolic leucine-zipper-like protein functioning in mature organs of soybean shoot against water-potential changes. (c) 2005 Elsevier B.V. All rights reserved., ELSEVIER SCIENCE BV
Gene, 2005年, ［査読有り］

Cloning of two cysteine proteinase genes, CysP1 and CysP2, from soybean cotyledons by cDNA representational difference analysis　　　　　　　　　　　　　　　
Ling J.Q; Kojima T; Shiraiwa M; Takahara H
Biochimica et Biophysica Acta, 2003年, ［査読有り］

Structural features of a wheat plastome as revealed by complete sequencing of chloroplast DNA
Ogihara Y.; Isono K.; Kojima T.; and 16 authors., Structural features of the wheat plastome were clarified by comparison of the complete sequence of wheat chloroplast DNA with those of rice and maize chloroplast genomes. The wheat plastome consists or a 134,545-bp circular molecule with 20,703-bp inverted repeats and the same gene content as the rice and maize plastomes. However, some structural divergence was found even in the coding regions of genes. These alterations are due to illegitimate recombination between two short direct repeats and/or replication slippage. Overall comparison of chloroplast DNAs among the three cereals indicated the presence of some hot-spot regions for length mutations. Whereas the region with clustered tRNA genes and that downstream of rbcL showed divergence in a species-specific manner, the deletion patterns of ORFs in the inverted-repeat regions and the borders between the inverted repeats and the small single-copy region support the notion that wheat and rice are related more closely to cacti other than to maize., SPRINGER-VERLAG
Molecular Genetics and Genomics, 2002年, ［査読有り］

Large-scale selection of lines with deletions in chromosome 1B in wheat and applications for fine deletion mapping
Tsujimoto H.; Yamada T.; Hasegawa K.; Usami N.; Kojima T.; Endo T.R.; Ogihara Y.; Sasakuma T., Terminal deletions of chromosome 1B in common wheat were selected on a large scale. The gametocidal gene of Aegilops cylindrica was used as the inducer of chromosome breakage. First, genes for endosperm storage proteins located on both arms of chromosome 1B were used as the selection markers. However, it was found that the chromosome breakage occurred during female gametogenesis, causing genotypic inconsistency between the embryo and endosperm. Thus, we isolated plants with terminal deletions in chromosome 1B by C-banding. Of 1327 plants examined, 128 showed aberrations in chromosome 1B: 47 in the short arm, 76 in the long arm, and 5 in both arms. The present deletions tended to have the breakpoint at more proximal regions than those produced previously by T.R. Endo and B.S. Gill. Using 33 deletion lines produced in this study and 34 lines previously produced, we mapped 39 RFLP loci and a nucleolar organizer region (NOR) on a specific region of chromosome 1B. The NOR was found to consist of two subregions with different repetitive units, which were termed NOR-B1d and NOR-B1p. Based on this fine deletion map and genotypic inconsistency between embryo and endosperm, the features of the gametocidal gene are discussed., NATL RESEARCH COUNCIL CANADA
Genome, 2001年, ［査読有り］

RAPD and ISSR fingerprints as useful genetic markers for analysis of genetic diversity, varietal identification, and phylogenetic relationships in peanut (Arachis hypogaea) cultivars and wild species
Raina S.N.; Rani V.; Kojima T.; Ogihara Y.; Singh K.P.; Devarumath R.M., Twenty-one random and 29 SSR primers were used to assess genetic variation and interrelationships among subspecies and botanical varieties of cultivated peanut, Arachis hypogaea (2n = 4x = 40), and phylogenetic relationships among cultivated peanut and wild species of the genus Arachis. In contrast with the previous generalization that peanut accessions lack genetic variation, both random and SSR primers revealed 42.7 and 54.4% polymorphism, respectively, among 220 and 124 genetic loci amplified from 13 accessions. Moreover, the dendrograms based on RAPD, ISSR, and RAPD + ISSR data precisely organized the five botanical varieties of the two subspecies into five clusters. One SSR primer was identified that could distinguish all the accessions analysed within a variety. Although the polymorphic index content varied from 0.1 to 0.5 for both ISSR and RAPD markers, primer index values were substantially higher for RAPD primers (0.35-4.65) than for SSR primers (0.35-1.73). It was possible to identify accessions, particularly those of divergent origins, by RAPD and (or) ISSR fingerprints. Based on these results, marker-based genetic improvement in A. hypogaea appears possible. None of the 486 RAPD and 330 ISSR amplification products were found to be commonly shared among 13 species of section Arachis and one species each of sections Heteranthae, Rhizomatosae, and Procumbentes. Dendrograms constructed from RAPD, ISSR, and RAPD + ISSR data showed overall similar topologies. They could be resolved into four groups corresponding to the species grouped in four taxonomic sections. The present results strongly support the view that Arachis monticola (2n = 4x = 40) and A. hypogaea are very closely related, and indicate that A. villosa and A. ipaensis are the diploid wild progenitors of these tetraploid species., CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
Genome, 2001年, ［査読有り］

Abundant DNA markers detected by PCR using simple sequence repeat primers without anchor sequences in soybean
Yang D.J.; Naito Y.; Ishikawa M.; Kojima T.; Tazuke A.; Niwa M., JAPANESE SOC BREEDING
Breeding Science, 2001年, ［査読有り］

Direct isolation of differentially expressed genes from a specific chromosome region of common wheat: application of the amplified fragment length polymorphism-based mRNA fingerprinting (AMF) method in combination with a deletion line of wheat
Kojima T.; Habu Y.; Iida S.; Ogihara Y., 筆頭著者, The amplified restriction fragment length polymorphism (AFLP)-based mRNA fingerprinting (AMF) method makes it possible systematically and conveniently to identify differentially expressed cDNAs with high reproducibility. We have applied the AMF method to the cloning of the Q gene of common wheat, which is located on the long arm of chromosome 5A and pleiotropically controls the spike morphology and the threshing character of seeds. Using the AMF method, we compared the fingerprints of mRNA samples extracted from the young spikes of Triticum aestivum cv. Chinese Spring (CS) carrying the Q gene to those of a chromosome deletion line of CS, namely, q5, which lacks 15% of 5AL including the Q gene. Approximately 12,200 fragments were produced after PCR with 256 primer combinations. Of these, 92 fragments were differentially expressed between CS and q5. Northern and Southern analyses showed that 16 fragments gave specific or relatively stronger transcript signals in CS, and these clones were present in single copy or in low copy numbers in the wheat genome. Four clones were genetically mapped to the region deleted in q5. Subsequently, one clone, pTaQ22, was mapped at the same locus as the Q gene, indicating that pTaQ22 corresponds to the Q gene or is tightly linked to it. DNA sequence data showed that pTaQ22 had no homology to any known genes, thus suggesting a novel function for this gene in flower morphogenesis. This AMF method might provide a straightforward method for isolating genes in the hexaploid background of common wheat., SPRINGER VERLAG
Molecular and General Genetics, 2000年, ［査読有り］

Chinese Spring wheat (Triticum aestivum L.) chloroplast genome: complete sequence and contig clones
Ogihara Y.; Isono K.; Kojima T. and 16 authors, Libraries of plasmid clones covering the entire chloroplast (cp) genome of the common wheat, Triticum aestivum cv. Chinese Spring were constructed and assembled into contig-clones. From these, we obtained the complete nucleotide sequence of wheat chloroplast DNA - a 134,540 bp circular DNA (DDBJ accession no. AB042240) containing four species of ribosomal RNA, 30 genes for 20 species of transfer RNA, and 71 protein coding genes. Additionally, we detected five unidentified open reading frames conserved among grasses. Plasmid clones are available on request., INT SOC PLANT MOLECULAR BIOLOGY
Plant Molecular Biology Reporter, 2000年, ［査読有り］

Genetic linkage map of ISSR and RAPD markers in Einkorn wheat in relation to that of markers
Kojima T.; Nagaoka T.; Noda K.; Ogihara Y., 筆頭著者, The potential of PCR-based markers for construction of a genetic linkage map in Einkorn wheat was investigated. From a comparison of polymorphisms between two Einkorn wheats, Triticum monococcum (Mn) and T. boeoticum (Bt), we obtained 49 polymorphic bands produced by 33 primers for inter-simple sequence repeat (ISSR) and 36 polymorphic bands shown by 25 combinations of random amplified polymorphic DNA (RAPD) primers for mapping in 66 individuals in the F-2 population. Although 44 ISSR fragments and 29 RAPD fragments statistically showed a 3:1 segregation ratio in the F-2 population, only 9 markers each of the ISSR and RAPD bands were able to be mapped on the RFLP linkage map of Einkorn wheat. ISSR markers were distributed throughout the chromosomes. The mapped positions of the ISSR markers seemed to be similar to those obtained by the RFLP markers. On the other hand, 4 of the 9 RAPD markers could map the RFLP marker-poor region on the short arm of 3A(m), suggesting a potential to map novel regions containing repetitive sequences. Comparisons of the genetic linkage map of Einkorn wheat to the linkage map and cytological map of common wheat revealed that the marker orders between the two maps of Einkorn wheat and common wheat coincided except for 4A, which harbors chromosome rearrangements specific for polyploid wheats, indicating a conservatism between the two genomes. Recombinations in Einkorn wheat chromosomes took place more frequently around the centromere and less at the distal part of chromosomes in comparison to those in common wheat. Nevertheless, recombinations even in Einkorn wheat chromosomes were strongly suppressed around the centromere. In fact, the markers located within 1 cM of the centromere were located almost in the central part of the chromosome arm., SPRINGER VERLAG
Theoretical and Applied Genetics, 1998年, ［査読有り］

High-resolution RFLP map of the long arm of chromosome 5A in wheats and its synteny among cereals
Kojima T.; Ogihara Y., 筆頭著者, A fine genetic map of the long arm of 5A in common wheat was generated by mapping 48 RFLP markers, in which 23 cDNAs were involved. That map was compared with the cytological map and a genetic map of the diploid obtained from a cross between Triticum monococcum and T. boeoticum. The marker distribution of the genetic map of common wheat almost correlated with the cytological distances, but some regions in which recombination was suppressed were found; these were around the centromere and in the region adjacent to the central C-banding region (L1.5). Promotion of recombination at the distal part of chromosome was not found. The marker order of the genetic map of common wheat was identical to that of the diploid, indicating that reciprocal translocation 4AL-5AL in wheat had taken place at the diploid level. Furthermore, this region was compared with the corresponding one of barley (5HL) and of rice (chromosome 9). Translocation was not detected in the barley chromosome, but paracentric inversion was found in comparison with the corresponding region of wheat. The translocation point in wheat seemed. to correspond to the inversion point of barley, suggesting a hot-spot region for chromosome breakage. Strong suppression of recombination was found in the central part of 5HL. About two-thirds of the proximal region of wheat 5AL revealed synteny with the corresponding region of rice, whereas there was no such relation in the more distal portion. Longer genetic distances in the central part of 5AL, in comparison with the corresponding region of rice, were found, indicating the presence of factors that affect recombination frequencies. The analyses suggest that the proximal part of chromosome arm 5AL has been relatively conserved, whereas the distal part has undergone more extensive rearrangements during evolution., GENETICS SOC JAPAN
Genes and Genetic Systems, 1998年, ［査読有り］

High-resolution RFLP mapping of the fertility restoration (Rf3) gene against Triticum timopheevi cytoplasm located on chromosome 1BS of common wheat
Kojima T.; Tsujimoto H.; Ogihara Y., 筆頭著者, Near-isogenic lines of alloplasmic wheat for the Rf3 gene controlling fertility restoration against the cytoplasm of Triticum timopheevi were developed by successive backcrossing of Chinese Spring to the F-1 plants between (timopheevi)-CS (male sterile) and T. spelta (carrying Rf3). The resultant three BC3F1 plants were self-pollinated so as to obtain 125 BC3F2 progenies. Using this population, we precisely mapped the Rf3 gene on the short arm of wheat chromosome 1B (1BS) of the RFLP linkage map. The Rf3 gene was localized at a position 1.2 cM and 2.6 cM distant from Xcdo388 and Xabc156, respectively. The genetic distances of Rf3 from Nor and Gli-BI were calculated to be 22.3 cM and 18.6 cM, respectively, supporting previous data. Estimation of the physical distance of the region suggets that the Rf3 gene resides within 500 kbp from the adjacent RFLP markers. The marker order of the genetic map corresponded to that of the cytological map, except for the position of Xbcd98. Comparison between genetic and cytological maps clearly shows that RFLP markers are unevenly distributed throughout the chromosome arm, and recombinations took place unequally in the chromosome arm., GENETICS SOC JAPAN
Genes and Genetic Systems, 1997年, ［査読有り］

Transcriptional regulation of peptidylargine deiminase expression in human keratinocytes
Ying S.; Dong S.; Kawada A.; Kojima T.; Chavanas S.; Méchin M-C.; Adoue V.; Serre G.; Simon M.; Takahara H.
Journal of Dermatological Science, 2009年, ［査読有り］, ［招待有り］

塩ストレス耐性に関与するダイズ酸性PR-5タンパク質の機能解析　　　　　　　　　　　　　　　
小島 俊雄、窪田 一乃、寺田 愛唯、會澤 由菜、立花 桃香、相馬 明佳、菊池 裕貴
日本農芸化学会2025年度大会, 2025年03月07日

塩ストレス応答・耐性機構におけるダイズPR-5タンパク質GmOLPaの機能解析　　　　　　　　　　　　　　　
相馬 明佳、竹下 聡香、菊池 裕貴、小島 俊雄
日本農芸化学会2023年度大会, 2023年03月14日

塩ストレス応答性を示すダイズPR-5タンパク質GmOLPaの分子学的特徴付け　　　　　　　　　　　　　　　
菊池 裕貴・小島 俊雄
第38回日本植物バイオテクノロジー学会, 2021年09月11日

形質転換シロイヌナズナを用いたダイズ塩ストレス応答遺伝子GmTDF-5の機能解析　　　　　　　　　　　　　　　
鈴木 佳奈、小川 佳祐、小島 俊雄
第38回日本植物バイオテクノロジー学会, 2021年09月11日

ダイズ耐塩性関連遺伝子GmTDF-5のプロモーター解析　　　　　　　　　　　　　　　
黒澤 まりな、牧野 果子、高木 ひかり、須藤 弘樹、野口 和斗、小島 俊雄
日本農芸化学会2021年度大会, 2021年03月20日

アルマジロリピートタンパク質をコードするタバコ機能未知遺伝子の構造と塩ストレス応答性　　　　　　　　　　　　　　　
後藤 里奈、小島 俊雄
第37回日本植物細胞分子生物学会, 2019年09月

ダイズ耐塩性に関わる機能未知タンパク質GmTDF-5のユビキチン－プロテアソームによる制御　　　　　　　　　　　　　　　
小川 佳祐、木村 愛、佐藤 光朗、小島 俊雄
日本農芸化学会2016年度大会, 2016年03月

ダイズ塩ストレス応答における機能未知遺伝子GmTDF-5の転写制御機構　　　　　　　　　　　　　　　
須藤 弘樹、鈴木 美緒、木村 愛、野口 和斗、小島 俊雄
日本農芸化学会2016年度大会, 2016年03月

植物の耐塩性に関与する機能未知タンパク質GmTDF-5のユビキチン-プロテアソームによる制御　　　　　　　　　　　　　　　
木村 愛、佐藤 光朗、高原 英成、小島 俊雄
日本農芸化学会2014年度大会, 2014年03月, 日本農芸化学会

ダイズの塩ストレス応答機構に関わる核-細胞質間輸送因子Importin αの探索　　　　　　　　　　　　　　　
川崎 裕史、佐川 智美、木村 愛、高原 英成、小島 俊雄
第31回日本植物細胞分子生物学会, 2013年09月

新規ダイズ塩ストレス応答遺伝子を導入したシロイヌナズナ培養細胞と植物体の耐塩性比較　　　　　　　　　　　　　　　
斎藤 祐一、高原 英成、小島 俊雄
日本農芸化学会2013年度大会, 2013年03月, 日本農芸化学会

ダイズの耐塩性を制御する機能未知アルマジロリピートタンパク質の塩ストレス応答性　　　　　　　　　　　　　　　
佐藤 光朗、右田 和琴、斎藤 祐一、野口 和斗、高原 英成、小島 俊雄
日本農芸化学会2013年度大会, 2013年03月, 日本農芸化学会

塩・乾燥ストレスに応答するミヤコグサARM-repeat Protein遺伝子の特徴付け　　　　　　　　　　　　　　　
野口和斗、斎藤祐一、梅崎修平、高原英成、小島俊雄
日本農芸化学会2012年度大会, 2012年03月

ミヤコグサ新規ARMタンパク質の遺伝子構造と水分ストレス応答性　　　　　　　　　　　　　　　
梅崎 修平、木下 美幸、山田 知恵、祝迫 まゆみ、野口 和斗、高原 英成、小島 俊雄
第28回日本植物細胞分子生物学会, 2010年09月

羽毛分解糸状菌から単離したケラチン誘導性遺伝子KI-2の分子学的特徴付け　　　　　　　　　　　　　　　
櫻井 雅詞、西舘 邦瑛、高原 英成、小島 俊雄
日本農芸化学会2009年度大会, 2009年03月

新規ダイズ遺伝子GmTDF-5を過剰発現する形質転換植物の耐塩性評価　　　　　　　　　　　　　　　
牧山 太樹、木下 美幸、山田 知恵、小島 俊雄、白岩 雅和、高原 英成
日本農芸化学会2008年度大会, 2008年03月

ダイズOxysterol-binding proteinに結合する脂質の探索　　　　　　　　　　　　　　　
北川 貴規、高橋 暢維、小島 俊雄、白岩 雅和、高原 英成
日本農芸化学会2008年度大会, 2008年03月

羽毛分解糸状菌に由来する新規遺伝子KI-1の羽毛ケラチン誘導性の解析　　　　　　　　　　　　　　　
吉田 藍、山本 慧、櫻井 雅詞、西舘 邦瑛、小島 俊雄、高原 英成
日本農芸化学会2008年度大会, 2008年03月

ダイズ塩ストレス応答性遺伝子GmTDF-5の過剰発現が及ぼす植物耐塩性への影響　　　　　　　　　　　　　　　
牧山太樹、高橋亜理沙、兼上明美、小島俊雄、白岩雅和、高原英成
第25回日本植物細胞分子生物学会, 2007年08月

羽毛分解糸状菌Chrysosporium keratinophilum 5M1株のケラチン誘導性遺伝子KI-1の構造解析　　　　　　　　　　　　　　　
西舘邦瑛、中屋俊樹、櫻井雅詞、小山洋一、林田治、楠畑雅、小島俊雄、高原英成
日本農芸化学会2007年度大会, 2007年03月

ケラチン誘導性遺伝子KI-1の組換え蛋白質発現系の構築とその機能解析　　　　　　　　　　　　　　　
小島俊雄、西舘邦瑛、菊池泉、吉田藍、佐野文子、高原英成
日本農芸化学会2007年度大会, 2007年03月

ダイズPR-5タンパク質群における遺伝子構造とストレス応答性の比較解析　　　　　　　　　　　　　　　
舘裕之、福田久美子、小島俊雄、白岩雅和、高原英成
日本農芸化学会2006年度大会, 2006年03月

塩ストレスに応答するダイズオキシステロール結合蛋白質の遺伝子単離と構造解析　　　　　　　　　　　　　　　
井上速見、高橋暢維、李東艶、小島俊雄、白岩雅和、高原英成
第28回日本分子生物学会, 2005年12月

ストレス/非ストレス環境におけるダイズオキシステロール結合蛋白質の遺伝子発現制御解析　　　　　　　　　　　　　　　
高橋暢維、井上速見、李東艶、小島俊雄、白岩雅和、高原英成
第28回日本分子生物学会, 2005年12月

塩ストレス応答における新規ダイズ遺伝子GmTDF-5の機能解析　　　　　　　　　　　　　　　
兼上明美、小島俊雄、白岩雅和、高原英成
第27回日本分子生物学会, 2004年12月

塩ストレスに対する新規ダイズ遺伝子GmTDF-5の転写制御機構　　　　　　　　　　　　　　　
三原迪子、大江秀志、小島俊雄、白岩雅和、高原英成
第27回日本分子生物学会, 2004年12月

ダイズPR-5タンパク質群の遺伝子構造と塩ストレス応答性の比較　　　　　　　　　　　　　　　
舘裕之、大西真理、小島俊雄、白岩雅和、高原英成
第22回日本植物細胞分子生物学会, 2004年08月

ダイズにおける塩ストレス誘導性遺伝子GmTDF-5の解析　　　　　　　　　　　　　　　
兼上明美、青木亜矢子、小島俊雄、白岩雅和、高原英成
日本農芸化学会2004年度大会, 2004年03月

ダイズにおける塩ストレス応答性β-グルコシダーゼ遺伝子の単離および遺伝子発現解析　　　　　　　　　　　　　　　
小椋真紀、青木亜矢子、小島俊雄、白岩雅和、高原英成
第26回日本分子生物学会, 2003年12月

ダイズプロリン代謝関連酵素遺伝子群の発現解析　　　　　　　　　　　　　　　
前田良彦、緒形沙百合、青木亜矢子、小島俊雄、白岩雅和、高原英成
第26回日本分子生物学会, 2003年12月

塩ストレスに応答するダイズ由来PR-5タンパク質、GmOLPの遺伝子解析　　　　　　　　　　　　　　　
小島俊雄、大西真理、白岩雅和、高原英成
第21回日本植物細胞分子生物学会, 2003年08月

CysP1 and CysP2, encoding two cysteine proteinases with C-terminal KDEL motif, are soybean cotyledon senescence-associsated genes　　　　　　　　　　　　　　　
Ling; J.Q.; Kojima; T.; Shiraiwa; M.; Takahara; H.
第25回日本分子生物学会, 2002年12月

ダイズオスモチン遺伝子のストレス応答性とその発現制御機構　　　　　　　　　　　　　　　
大西真理、小島俊雄、白岩雅和、高原英成
第25回日本分子生物学会, 2002年12月

ダイズOSBP遺伝子の単離とその遺伝子解析　　　　　　　　　　　　　　　
李東艶、青木亜矢子、小島俊雄、白岩雅和、高原英成
第25回日本分子生物学会, 2002年12月

ダイズにおける塩ストレス応答性遺伝子群のcDNA-AFLP解析　　　　　　　　　　　　　　　
小島俊雄、青木亜矢子、白岩雅和、高原英成
第23回種子生理生化学研究会, 2002年11月

ダイズ塩ストレス誘導制オスモチン遺伝子の単離とストレス応答性解析　　　　　　　　　　　　　　　
大西真理、小島俊雄、白岩雅和、高原英成
第23回種子生理生化学研究会, 2002年11月

Molecular cloning of two soybean cysteine proteinase genes in senescing cotyledon by cDNA representational difference analysis　　　　　　　　　　　　　　　
Ling; J.Q.; Kojima; T.; Shiraiwa; M.; Takahara; H.
日本農芸化学会2002年度大会, 2002年03月

ダイズにおける塩ストレス誘導性オスモチン遺伝子の単離と発現解析　　　　　　　　　　　　　　　
大西真理、青木亜矢子、小島俊雄、白岩雅和、高原英成
日本農芸化学会2002年度大会, 2002年03月

塩ストレスに対するダイズプロリン生合成関連酵素の遺伝子解析　　　　　　　　　　　　　　　
小島俊雄、緒形沙百合、大西真理、前田良彦、青木亜矢子、白岩雅和、高原英成
第22回種子生理生化学研究会, 2001年11月

ダイズにおける塩誘導性遺伝子群の発現解析　　　　　　　　　　　　　　　
青木亜矢子、小島俊雄、白岩雅和、高原英成
第23回日本分子生物学会, 2000年12月

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