Annual Report 2006

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and time course-dependent manner with minimum expression at 1 µM GA3 and maximum expression at 50 µM GA3 starting from 1 h and peaked at 24 h after GA3 treatment. expression was up regulated by GA3 at transcript level while no significant effect was observed for other hormones. OsGAE1 was expressed in with N-terminal His6 tag and the recombinant protein migrated at 38 kDa, slightly larger than the predicted 29 kDa, during SDS-polyacrylamide gel electrophoresis. Anti-OsGAE 1 antibodies immunoreacted with a protein of 40 kDa in rice leaf sheath. expressed mainly in growing leaf sheath and callus compared to leaf and root. hybridization and promoter analysis revealed that expressed in shoot apex meristem and young primary leaves (Fig. 3). Northern blot, Western blot and GUS activities revealed that OsGAE1 transcript and translation are up regulated by GA3. Transgenic rice expressing in antisense orientation exhibited severely affected vegetative and reproductive growth (Fig. 4). The transgenic plants were 55 to 70 % short compared to control. These results suggest that OsGAE1 is differentially expressed in rice leaf sheath in relation to GA3 and it encodes a functional protein which is involved in GA regulated growth and development of rice. Fig. 4 Phenotype of transgenic rice constitutively expressing in antisense direction a) The transgenic plants were grown in an isolated greenhouse and photographed two months after transfer to soil. b) Antisense transgenic plants exhibited stem bifurcation usually at 2nd or 3rd node. c) Spikes of antisense transgenic plants remained in the leaf sheath of flag leaf.
B iochemistry Department Structural biology X-ray crystallographic analysis of proteins Crystal structure studies of several biologically important proteins have been carried out. α-Galactosidases catalyze the hydrolysis of α-linked galactosyl residues from galacto-oligosaccharides and polymeric galacto- (gluco)mannans. The crystal structure of α-galactosidase I was determined at 1.6 resolution (Fig. 1). The structure consisted of a catalytic domain comprising a (β/α)8-barrel structure and a C- terminal domain made up of eight β-strands containing a Greek key motif. Owing to the high resolution X-ray data, four carbohydrate chains were observed in one α-galactosidase I molecule and their structures were identified to be high mannose type. α-Galactosidase I seemed to form a tetramer around the crystallographic four-fold axis. The crystal structure of elapid snake toxins pseudechetoxin (PsTx) and pseudecin (Pdc) have been determined at around 2 resolution. These proteins belong to the cysteine-rich secretory protein family isolated from the snake venom and target cyclic nucleotide-gated ion channels. The structure consisted of an N-terminal domain that has a fold similar to the group 1 plant pathogenesisrelated proteins and a cysteine-rich C-terminal domain. The multidomain strucutre seemed to play an important role to recognize the target proteins. 3D-structure of barnacle cement protein, -20k Structure determination by X-ray crystallography and/or NMR spectroscopy is the powerful tool for research of proteins with unknown functions because protein function is strictly regulated by three-dimensional (3D) structure. Barnacle cement proteins, which are secreted for underwater adhesion, were recently isolated and cloned. The molecular functions of these proteins in adhesion are not to be established. We have determined the 3Dstructure of one of these proteins, cp-20k, in solution by NMR spectroscopy in order to obtain insight into its biological function. cp-20k contains 32 cysteine residues. They are assembled in regular repetitive positions in the primary structure, leading to Fig. 1 The ribbon model of the crystal structure of α-galactosidase I Two catalytic residues, disulfide bridges and the sugar chains were shown as black balland-stick drawings in red, yellow and gray color, respectively. Fig.2 Structure of -20k Six homologous units are numbered, and the boundaries of each repeats are marked by dotted lines. Cystines are shown in yellow sticks.
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2006
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Teruo Ishige, President The Nationa
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List of Publication Original paper
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Topics of Research in This Year Pos
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studies. To construct a fully satur
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Fig. 2 Geographical distribution of
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(ABR0015, ABR0257, ABR0417, ABR0495
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Mulberry latex defense mulberry tre
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Fig. 2 Growth inhibitory effects of
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Fig. 3 Change of the levels of luci
Page 21 and 22: Next, we examined the effect of tra
Page 23 and 24: G enome Research Department Introdu
Page 25 and 26: The Rice Annotation Project Databas
Page 27 and 28: industrial and agricultural concern
Page 29 and 30: One of the main problems involving
Page 31 and 32: Fig. 3 Mapping of QTL for rachis in
Page 33 and 34: two genes, XOO2263 and XOO4134 cont
Page 35 and 36: G enebank Genebank Project The NIAS
Page 37 and 38: using dormant winter buds. Also, we
Page 39 and 40: Insect and Animal Sciences Division
Page 41 and 42: Expression patterns of two novel ge
Page 43 and 44: Fig. 5 PGC and EBC fused cells PGCs
Page 45 and 46: cells. The epidermal cells maintain
Page 47 and 48: Animal and cell culture models for
Page 49 and 50: terminus, respectively. A signal se
Page 51 and 52: Fig. 2 Trehalose content of body pa
Page 53 and 54: Fig. 6 Birth weight of 11 adult SNT
Page 55 and 56: The role of glucose as a metabolic
Page 57 and 58: analysis using microsatellite marke
Page 59 and 60: Fig. 2 Recorded Electromyograms (le
Page 61 and 62: Table 1 Menu of Commercial silkworm
Page 63 and 64: Fig. 3 Expression of GFP gene in th
Page 65 and 66: also surveyed in the genetic stocks
Page 67 and 68: esearch were located in or near gen
Page 69 and 70: elatives results in severe reductio
Page 71: and circadian clocks may control th
Page 75 and 76: Proteomic analysis Based on the pro
Page 77 and 78: photoautotrophic medium (Fig. 1B).
Page 79 and 80: Fig. 4 Blast resistance of OsWRKY45
Page 81 and 82: phosphatase activity by SIPK depend
Page 83 and 84: the endosperm in co-operation with
Page 85 and 86: isolated genes in transgenic Anal
Page 87 and 88: Fig. 1 Gene structures and mutation
Page 89 and 90: cytochimera among the mulberries (
Page 91 and 92: 15 G. Hariraj, Kinoshita Haruo, T.
Page 93 and 94: 49 Kameda Tsunenori, Teramoto Hidet
Page 95 and 96: 85 Matsuyama Shuichi, Ohkura Satosh
Page 97 and 98: 120 Ozawa Kenjiro, Kawahigashi Hiro
Page 99 and 100: 155 Taguchi-Shiobara Fumio, Yamamot
Page 101 and 102: 192 Ueno Osamu, Agarie Sakae, (2005
Page 103 and 104: 8 Murakami Ritsuko, Koyama Akio, Sh
Page 105 and 106: 20 Todoroki Junichi, Kaneko Hiroyuk
Page 107 and 108: Executive Members and Research Staf
Page 109 and 110: Developmental Biology Department Di
Page 111 and 112: Research Leader Hayasaka Shoji Rese
Page 113 and 114: Mutation Genetics Laboratory Head N
Page 115 and 116: thousands of yen TOTAL BUDGET OPERA
Page 117: Annual Report 2006 (Apr. 2005Mar. 2
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Annual Report 2006

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