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Construction of the Seed-Coat cDNA Microarray and Screening of Differentially Expressed Genes in Barley

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 695-700页

作者： Jin-Song PANG*, Meng-Yuan HE, and Bao LIU Laboratory of Molecular Epigenetics, institute of genetics and cytology, northeast normal university, changchun 13002., china Laboratory of Molecular Epigenetics Institute of Genetics and Cytology Northeast Normal University

Some barley mutants can synthesize neither anthocyanins nor proanthocyanidins in the seed coat, which is related to several genes in locus Ant13, but the exact model of action remains unknown. We used the cDNA microarray technology with barley transcription-deficient mutant (ant13-152. that does not synthesize proanthocyanidins as the tester, and its wild type genotype (Triumph) as the driver, to study this question. Six-thousand and forty-eight clones from the wild type Morex testa+pericarp cDNA library were amplified using PCR, and the DNA fragments were spotted on commercial amino-modified glass slide as microarray. The mRNAs from the developing seed coat (8–15 days) of both the mutant and the wild-type barley plants were isolated, and labeled respectively with Cy3-dUTP and Cy5-dUTP when reversely trans- cribed to cDNAs. The labeled cDNAs were used as probes, mixed at the same molar concentration, and hybridized with the DNA fragments on the slide. Seventy clones exhibiting marked differential expression (ratio>4) were identified from the microarray. All the 2. cDNA clones that showed an over-expression in wild type in comparison to the mutant ant13-152.were sequenced. It was found that most of these over- expressing clones were transcription/translation and hordein-associated genes. These results have laid a solid material basis for further elucidation of the metabolic pathway in proanthocyanidin synthesis in barley and likely other plants.

关键词： barley seed coat proanthocyanidin cDNA microarray

Immunolcocalization of actin in intact and DNA-and histone-depleted nuclei and chromosomes of allium cepa

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Cell Research 1998年第1期8卷 51-62页

作者： WAN LI HONG MIAO XING (institute of genetics and {2., northeast normal university, changchun 13002., china) institute of genetics and {2. Northeast Normal University China

The presence of actin in eukaryotic nuclei and chromosomes, and especially in higher plant nuclei and chromosomes, has not been well established. We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes, indicating that actin is present in the nuclei and chromosomes of the higher plant. We labeled sections of the meristematic cells of A. cepa with immunogold technique, gold particles were found over the whole nuclei and a number of gold particles were concentrated in condensed chromatin and nucleoli, confirming the results of the immunofluoresence observations. We treated the nuclei and chromosomes of *** with DNase I and 2. NaCl and obtained DNA- and histone-depleted nuclei and chromosomes. Indirect immunofluorescence tests showed that the DNA- and histonedepleted nuclei and chromosomes reacted positively with the anti-actin antibodies. These results demonstrate that actin exists not only in intact nuclei and chromosomes,but also in DNA- and histone-depleted nuclei and chrmosomes of the plant. In addition, our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A. cepa.

关键词： Actin Allium cepa chromosomes nuclei

Tropomyosin is localized in the nuclear matrix and chromosome scaffold of physarum polycephalum

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Cell Research 1999年第1期9卷 61-69页

作者： ZENG XIAN LU MING DA JIAO MIAO XING XIAO GUANG WANG SHUI HAO(institute of genetics and {2., northeast normal university, changchun 13002., china)(Department of Biology, changchun normal College,changchun 130032. china) instituteofgeneticsand{2. NortheastNormalUniversityChangchunChina.

The nuclei and chromosomes were isolated from plasmodia of Physarum polycephalum. The nuclear matrir and chromosome scaffold were obtained after the DNA and most of the proteins were extracted with DNase I and 2.M NaCl. SDS-PAGE analyses revealed that the nuclear matrir and chromosome scaffold contained a 37 kD polypeptide which is equivalent to tropomyosin in molecular weight. Immunofluorescence observations upon slide preparations labeled with anti-tropomyosin antibody showed that the nuclear matrix and chromosome scaffold emanated bright fluorescence, suggesting the presence of the antigen in *** results confirmed the presence of tropomyosin in the nuclear matrix and chromosome scaffold. Immunoelectron microscopic obserwtions further demonstrated that tropomyosin was dispersively distributed in the interphase nuclei and metaphase chromosomes.

关键词： Chromosome scaffold nuclear matrix Physarum polycephalum tropomyosin

A PSTAIRE CDK-like protein localizes in nuclei and cytoplasm of Physarum polycephalum and functions in the mitosis

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Cell Research 2004年第2期14卷 169-175页

作者： Gui Ying LI~(1,2., Miao XING~(1,*), Bo HU~(1,3) ~1College of Life Science, Shenzhen Key Laboratory of Microbial Gene Engineering, Shenzhen university, Shenzhen 518060, china. ~2.epartment of Immunology, School of Basic Medical Sciences, Jilin university, changchun 13002., china ~3institute of genetics and cytology, northeast normal university, changchun 13002., china. College of Life Science Shenzhen Key Laboratory of Microbial Gene Engineering Shenzhen University Shenzhen 518060 China.

CDKs play key roles in controlling cell cycle progression in all eukaryotes. In plants, multiple CDKs are present, among which the best characterized CDKs are PSTAIRE CDKs. In this study, we carried out Western blot, immunoelectron microscopy and antibody treatment with an anti-PSTAIRE monoclonal antibody to explore the subcellular localization and functions of PSTAIRE CDKs in Physarum polycephalum. The results of Western blot and immunoelectron microscopy showed that in P. polycephalum, a PSTAIRE CDK-like protein was 34 kD in molecular weight and located in both nuclei and cytoplasm. In nuclei, the protein was mainly associated with chromosomes and nucleoli. The expression of the PSTAIRE CDK-like protein in both the plasmodia and nuclei showed little fluctuation through the whole cell cycle. When treated with an anti-PSTAIRE monoclonal antibody at early S phase, the cells were arrested in S phase, and the mitotic onset of P. polycephalum was blocked for about 1 h when treated at early G2.phase. Our dat

关键词： PSTAIRE CDK-like protein Physarum polycephalum mitosis immunoelectron microscopy.

Trichostatin A Extends the Lifespan of Drosophila melanogaster by Elevating hsp2. Expression

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 618-622页

作者： Dan TAO#, Jun LU#, Hui SUN, Yan-Mei ZHAO, Zhi-Gen YUAN, Xiao-Xue LI, and Bai-Qu HUANG* institute of genetics and {2., northeast normal university, changchun 13002., china institute of genetics and {2. Northeast Normal UniversityChangchun 130024China

The level of acetylation of histones in nucleosomes is related to the longevity of yeast and animals. However, the mechanisms by which acetylation and deacetylation affect longevity remain unclear. In present study, we investigated the influence of histone acetylation modification on the expression of hsp2. gene and the lifespan in Drosophila melanogaster using histone deacetylase (HDAC) inhibitor Trichostatin A (TSA). The results showed that TSA could extend the lifespan of Drosophila melanogaster. Furthermore, TSA significantly promoted the hsp2. gene transcription, and affected the chromatin morphology at the locus of hsp2. gene along the polytene chromosome. Present data implicate that TSA may affect the lifespan of Drosophila through changing the level of histone acetylation and influencing the expression of hsp2. gene that is related to aging.

关键词： histone acetylation histone deacetylase inhibitor Drosophila melanogaster hsp2. lifespan

Structural components of the nuclear body in nuclei of Allium cepa cells

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Cell Research 2001年第1期11卷 68-73页

作者： TAO WEI, CHANG HUI YAN, TAO CAI, SHUI HAO, ZHONG HE ZHAI (College of Life Sciences, Peking university, Beijing 100871, china) ({2. of genetics and cytology, northeast normal university, changchun Jilin 13002., china) 1College of Life Sciences Peking University Beijing 100871 China 1College of Life Sciences Peking University Beijing 100871 China 1College of Life Sciences Peking University Beijing 100871 China 2.0} of genetics and {2. Northeast Normal University Changchun Jilin 130024 China 1College of Life Sciences Peking University Beijing 100871 China

Nuclear bodies have long been noted in interphase nuclei of plant cells, but their structural component, origin and fUnction are still unclear by now. The present work showed in onion cells the nuclear bodies appeared as a spherical structure about 0.3 to 0.8 μm in diameter. They possibly were formed in nucleolus and subsequently released, and entered into nucleoplasm. Observation through cytochemical staining method at the ultrastructural level codeined that nuclear bodies consisted of ribonucleoproteins (RNPs) and silver-stainable proteins. Immunocytochemical results revealed that nuclear bodies contained no DNA and ribosomal gene transcription factor (UBF). Based on these data, we suggested that nuclear bodies are not related to the ribosome or other gene transcription activities, instead they may act as subnuclear structures for RNPs transport from nucleolus to cytoplasm, and may also be involved in splicing of pre-mRNAs.

关键词： Nuclear body structural component ribonucleoproteins nucleolus Allium cepa.

Preparation of Polyclonal Antibodies of Rubisco Large and Small Subunits and Their Application in the Functional Analysis of the Genes

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 644-648页

作者： Peng-Da MA1, Tian-Cheng LU1, Xiao-Fu ZHOU1,2. Xiao-Juan ZHU1, and Xing-Zhi WANG1*1The institute of genetics and {2., northeast normal university, changchun 13002., china 2.he institute of Gene Technology, Jilin normal university, Siping 136000, china The institute of genetics and {2. Northeast Normal University The institute of Gene Technology Jilin Normal University

Spinach Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) large (rbcL) and small (rbcS) subunits were separated by SDS-PAGE, and protein amount and purity were determined by Bradford assay. Polyclonal antibodies against rbcL and rbcS subunit were generated in female BALB/c mice and had no cross-reaction with each other. A total of 81 μg antigens were used and 0.3 ml anti-sera with titer of 1:5000 were yielded. The antibodies were also applicable to study rbcL and rbcS in tobacco plant Nicotiana benthamiana. Potato virus X vector pGR107 induced silencing of rbcS gene by Agrobacterium in Nicotiana benthamiana was performed. The expression level of rbcL and rbcS was lower in rbcS silenced plants than that in control plants as detected by the corresponding antibodies. This implied that the expression of rbcL was regulated by rbcS.

关键词： ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) rbcL rbcS polyclonal antibody virus-induced gene silencing (VIGS)

Alterations in DNA methylation and genome structure in two rice mutant lines induced by high pressure

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Science china(Life Sciences) 2006年第2期49卷 97-104页

作者： SHEN Sile1,3, WANG Zhenwei3, SHAN Xiaohui2. WANG Hua3, LI Ling3, LIN Xuyun2. LONG Likun2. WENG Kenan4, LIU Bao2.& ZOU Guangtian1 1. National Laboratory of Superhard Materials, Jilin university, changchun 130012.{2. 2. institute of genetics and cytology, northeast normal university, changchun 13002.,{2. 3. College of Life Science, Jilin university, changchun 130012.{2. 4. Guangzhou institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640,{2. 1. National Laboratory of Superhard Materials Jilin University Changchun 130012 China 3. College of Life Science Jilin University Changchun 130012 China 2. institute of genetics and cytology Northeast Normal University Changchun 130024 China 4. Guangzhou institute of Geochemistry Chinese Academy of Sciences Guangzhou 510640 China

By using high-pressure treatment, two mutant lines were obtained from agenetically stable japonica rice cultivar Bijing38. Genomic DNA of the mutant lines, together withthe original line (Bijing38), was either undigested or digested by Hpa ⅡIMspⅠ, and then subjectedto molecular analysis using two markers, ISSR and RAPD. Results indicated that changes in the PCRamplification profiles of both markers are apparent in the two mutant lines compared with theoriginal rice cultivar, suggesting that there had been both sequence changes and DNA methylationmodifications in the mutant lines. Southern blot analysis using diverse sequences, including twocellular genes (S2.and S3), a set of retrotransposons (Osr7, Osr36, Tos19 and more), and a MITEtransposon family (mPing and Pong), confirmed the results, and indicated that changes in DNAmethylation pattern, genomic structure, and possible activation of some transposons indeed occurredin the mutant lines. Moreover, these changes are stably maintained through selfed generations and indifferent organs. Thus, our results indicate that it is possible to obtain stable mutants in riceby high pressure treatments, and the molecular basis of the mutants may include both genetic andepigenetic changes. Therefore, high hydrostatic pressure seems a promising approach for plantmutagenesis.

关键词： DNA methylation, genomic changes, high pressure, mutagenesis, rice.

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A new definition of restricted Lie superalgebras

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Chinese Science Bulletin 2000年第4期45卷 316-321页

作者： WANG Ying & ZHANG Yongzheng1. Department of Mathematics, Harbin institute Technology, Harbin 150001, china 2. Department of Mathematics,{2., changchun 13002., china Department of Mathematics Harbin Institute Technology Harbin China Department of Mathematics Northeast Normal University Changchun China

A new definition of restricted Lie superalgebras has been given; some important results on the representation theory of restricted Lie superalgebra have been obtained.

关键词： restricted Lie superalgebra p-mapping p-semilinear mapping α-representation.

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