Molecular cloning of a chloroplastic proteinassociated with both the envelope and thylakoid membranes

Chloroplasts consist of six morphologically distinct compartments. Each compartment has a specific set of polypeptides that perform distinct biochemical functions. We report here the identification of a membrane-associated protein with a novel localization. This protein was synthesized as a 37 kDa precursor and was processed to a mature protein of 30 kDa after being imported into isolated pea chloroplasts. Fractionation of chloroplasts showed that the 30 kDa mature protein was associated with both of the envelope membranes as well as with thylakoid membranes. Immunocyto-chemical localization of the 30 kDa protein revealed that the protein occurred in clusters in the vicinity of both the envelope and the thylakoid. Possible functions of this 30 kDa protein, inferred from its novel localization pattern, are discussed.Abbreviations CAB light-harvesting chlorophyll a/b-binding protein of photosystem II - prCAB precursor protein to CAB - SS small subunit of ribulose-1,5-bisphosphate carboxylase - prSS precursor protein to SS - RCF relative centrifugation force     

Anatomy of the stimulative sequences flanking the ARS consensus sequence of chromosome VI in Saccharomyces cerevisiae

We have analyzed the relationship between autonomously replicating sequence (ARS) structure and function for three ARS (ARS605, ARS607 and ARS609) from chromosome VI of Saccharomyces cerevisiae by systematic XhoI-linker mutation in the ARS consensus sequence (ACS) and flanking sequences. All mutations that encroached upon the ACS destroyed ARS activity. DNA sequences stimulative for ARS function were identified on either side of the ACS of ARS605 and only on the 3'-side of the ACS of ARS607. In ARS609, however, no such stimulative sequences were observed. Base substitutions complementary to the wild-type sequence of those stimulative regions, in ARS605 and ARS607, that did not change the AG of unwinding nor affected ARS activity suggests that these regions have, at least, a function as DNA-unwinding elements (DUE). ARS605, ARS607 and ARS609 DNA are of low AG value and showed hypersensitivity to single-strand-specific nuclease when inserted in negatively supercoiled plasmid. Linker mutations inhibitory for ARS activity (5L11 and 7L14) also caused significant changes in local nucleotide (nt) sensitivity within the ACS and its adjoining regions. Complementary base substitutions, however, did not affect these changes in local nt sensitivity. These results imply that the stimulative regions flanking the ACS are necessary to produce an optimum conformation around the ACS which may be important for full ARS activity.     

A Similar Expression Pattern of Adhesion Molecules between Intermediate TCR Cells in the Liver and Intraepithelial Lymphocytes in the Intestine

Two major populations of extrathymically differentiated T cells exist in the liver and intestine. Such T cells in the liver have TCR of intermediate intensity (i.e., intermediate TCR cells) and constitutively express IL-2 receptor β-chain (IL-2Rβ), whereas those in the intestine, especially intraepithelial lymphocytes, have TCR of bright intensity, consisting of a mixture of IL-2Rβ⁺ and IL-2Rβ^–. All mature thymocytes and thymus-derived T cells seen in the peripheral immune organs are TCR-bright⁺IL-2Rβ^– under resting conditions. When the expression pattern of adhesion molecules, including CD44, L-selectin, LFA-1 and ICAM-1, was compared among these T-cell populations, they displayed quite unique patterns of expression. All extrathymic T cells in the liver, intestine, and even other organs were CD44⁺L-selectin^– LFA-1⁺⁺ICAM-1⁺, whereas thymocytes and thymus-derived T cells were CD44^– L-selectin⁺LFA-1⁺ICAM-1^–. This inverted expression of adhesion molecules between extrathymic T cells and thymus-derived T cells might be associated with their unique tissue-localization.     

The number of species occurring in a sample of a biotic community and its connections with species-abundance relationship and spatial distribution

Connections among species-abundance (i-m _i ), species-frequency (i-F _i ), and species-sample size (S _n -n) relationships were examined on the basis of the mapping data of a natural forest in Thailand. The spatial distribution of individual trees without any discrimination of species was nearly random. Provided that the spatial distribution of each species was random, thei-m _i and thei-F _i relationship was reconstructed from each other in terms of the total number of species (S) and the total number of individuals (N) in the data. The number of species (S _n ) in a subsample consisting ofn individuals was then obtained from thei-F _i relationship. Logarithm ofS _n increased with logn and showed a convex curve through the origin. The values of diversity indices based onN andS(orn andS _n ) were affected by sample size. These trends were further examined on the basis of 944 data sets of biotic communities and three mathematical models of anS-N relationship. The properties of species-area relation were discussed in the light of these results.     

Synthesis of sulfated derivatives of curdlan and their anti-HIV activity

Sulfopropyl curdlan was synthesized, its structure was determined, and the anti-HIV activity was compared with that of standard curdlan sulfates obtained with piperidine N-sulfonic acid in dimethyl sulfoxide. It was shown that sulfopropyl curdlan exhibits weaker anti-HIV activity than curdlan sulfate. Curdlan sulfates were synthesized with a SO₃-pyridine complex in a heterogeneous phase. It was shown from ¹³C-NMR spectra of acetylated curdlan sulfates that they had a different substituent distribution from standard curdlan sulfate. The cytotoxicity of the curdlan sulfates was attributed to their heterogeneous structure.     

Cloning of the blasticidin S deaminase gene (BSD) from Aspergillus terreus and its use as a selectable marker for Schizosaccharomyces pombe and Pyricularia oryzae

Aspergillus terreus produces a unique enzyme, blasticidin S deaminase, which catalyzes the deamination of blasticidin S (BS), and in consequence confers high resistance to the antibiotic. A cDNA clone derived from the structural gene for BS deaminase (BSD) was isolated by transforming Escherichia coli with an Aspergillus cDNA expression library and directly selecting for the ability to grow in the presence of the antibiotic. The complete nucleotide sequene of BSD was determined and proved to contain an open reading frame of 393 bp, encoding a polypeptide of 130 amino acids. Comparison of its nulceotide sequence with that of bsr, the BS deaminase gene isolated from Bacillus cereus, indicated no homology and a large difference in codon usage. The activity of BSD expressed in E. coli was easily quantified by an assay based on spectrophotometric recording. The BSD gene was placed in a shuttle vector for Schizosaccharomyces pombe, downstream of the SV40 early region promoter, and this allowed direct selection with BS at high frequency, following transformation into the yeast. The BSD gene was also employed as a selectable marker for Pyricularia oryzae, which could not be transformed to BS resistance by bsr. These results promise that the BSD gene will be useful as a new dominant selectable marker for eukaryotes.     

金花茶花药愈伤组织的体细胞减数分裂

金花茶(Camellia petelotii)的小孢子单核期花药,经培养和继代培养6个月后的愈伤组织中,发现有少量体细胞进行减数分裂。在减数第一和第二次分裂中,同源染色体的配对和分离基本正常,最后形成四分体。该愈伤组织经石蜡切片和压片观察,发现其主要由大量的液化细胞和贮藏细胞所组成,此外,还有少部分分生细胞。没有发现进一步的分化。其染色体数目,2n=30者占71.7%其余则为非整倍体。     

Involvement of Collagen Synthesis in Tissue Reconstitution by Dissociated Sponge Cells

Cells dissociated from the sponge Haliclona permollis reconstituted a new body by the following three consecutive processes: (1) Reaggregation, cells aggregate as a spheroidal mass. (2) Spreading, aggregates flatten and spread. (3) Reconstruction, aggregates coalesce into a reconstituted body. Cells in the process of reconstitution incorporated ¹⁴C-proline into proteins, converting 17% of it to hydroxyproline and synthesize collagen molecules during this process. Inhibitors of collagen biosynthesis did not affect the reaggregation, but caused incomplete morphogenesis in the processes of spreading and reconstruction: cycloheximide and 2,2'-dipyridyl inhibited spreading while 3-aminopropionitrile induced incomplete reconstruction. These findings suggested that spreading and reconstruction, but not reaggregation require both synthesis and cross-linking of collagens. Three polypeptides with molecular weights of 58 K, 160 K and 180 K were identified in sponge cells as collagens by immunoblot analysis with antibodies against sea urchin collagen and studies on susceptibilities to collagenase and pepsin. The 58 KDa polypeptide appeared in reconstituted bodies but not in dissociated cells, suggesting its importance in tissue reconstitution by dissociated cells.     

The development of emotion regulation ability between the ages of 13 and 18 months in Japanese infants during the strange situation as a function of mothers' styles of emotion expression

The purposes of this study were two-fold. First, it compared Japanese infants' (N=129) abilities to regulate emotions at 13 months and 18 months of age, using the Strange Situation procedure. Second, it examined the relationship between the development of emotion regulation and the mother's emotion expression style as assessed by the Emotion Expression Style Questionnaire (EESQ). The total number of subjects who successfully completed all 8 episodes of the Strange Situation procedure increased significantly during the aging between 13 and 18 months of age, indicating that as a group these infants increased their ability to cope with stressful situations. However, infants who had mothers with negative emotion expression styles did not show greater capacity for emotion regulation at 18 months. These findings suggest that the development of emotion regulation is mediated by the mother's emotion expression style.     

Light-induced phase-shifting of the peripheral circadian oscillator in the hearts of food-deprived mice.

In the present study, we investigated the effect of fasting on photoentrainment of the peripheral circadian oscillator in the mammalian heart. Northern blotting showed that a single light pulse applied at an appropriate time in constant darkness, caused obvious phase-shifting in the circadian expression rhythm of the mammalian clock gene Period2 (mPer2) even in the hearts of food-deprived mice. Fasting did not significantly affect either the phase or the light-induced phase-shifts of the mPer2 rhythm. Although several studies of temporal feeding restriction have indicated that feeding is the dominant timing cue for mammalian peripheral oscillators, our findings suggest that feeding is not essential for mammals to induce phase resetting of the circadian oscillator in the heart.