Weisheng Wu

Shanghai Academy of Public Measurement, Shanghai, Shanghai Shi, China

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Publications (16)31.94 Total impact

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    ABSTRACT: Ginkgo biloba contains terpene triclactones of high pharmaceutical value such as ginkgolides. 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate (HMBPP) reductase (HDR) is proved to be the terminal-acting enzyme in the plastid MEP pathway which provides isoprenoid precursors for the biosynthesis of ginkgolides. The full-length cDNA encoding HDR, designated as GbHDR (Genbank Accession Number DQ364231), was isolated for the first time from G. biloba by RACE method. GbHDR contained a 1,422-bp open reading frame encoding 474 amino acids. The deduced GbHDR protein, showing high identity to HDRs of other plant species, was predicted to possess a chloroplast transit peptide at the N-terminal and four conserved cysteine residues. Two-dimensional structural analysis showed that GbHDR had a similar secondary structure with HDR from Arabidopsis thaliana. Southern blot analysis indicated that GbHDR belonged to a small gene family. Transcription pattern analysis revealed that GbHDR had high transcription in roots, and low in leaves and stems. The cloning of GbHDR gene will enable us to further understand the role of GbHDR involved in terpene triclatones biosynthetic pathway in G. biloba at molecular level.
    Molecular Biology Reports 06/2007; 35(3):413-20. · 2.51 Impact Factor
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    ABSTRACT: Pinellia pedatisecta agglutinin (PPA)is a very basic protein that accumulates in the tuber of P.pedatisecta .PPA is a hetero-tetramer protein of 40 kDa,composed of two polypeptide chains A (about 12 kDa)and two polypeptides chains B (about 12 kDa).The full-length cDNA of PPA was cloned from P.pedatisecta using SMART RACE-PCR technology; it was 1146 bp and contained a 771 bp open reading frame (ORF)encoding a lectin precursor of 256 amino acid residues with a 24 amino acid signal peptide.The PPA precursor contained 3 mannose-binding sites (QXDXNXVXY) and two conserved domains of 43% identity,PPA-DOM 1 (polypeptides A)and PPA-DOM 2 (polypeptides B).PPA shared varying identities,ranging from 40% to 85%,with mannose-binding lectins from other species of plant families such as Araceae, Alliaceae, Iridaceae, Liliaceae, Amaryllidaceae and Bromeliaceae. Southern blot analysis indicated that ppa belonged to a multi-copy gene family. Expression pattern analysis revealed that ppa expressed in most tested tissues, with high expression being found in spadix,spathe and tuber.Cloning of the ppa gene not only provides a basis for further investigation of its structure,expression and regulatory mechanism,but also enables us to test its potential role in controlling pests and fungal diseases by transferring the gene into plants in the future.
    Journal of Biosciences 04/2007; 32(2):241-9. · 1.76 Impact Factor
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    ABSTRACT: Tubers of Pinellia ternata are one of the well known traditional Chinese medicines. According to the Chinese Pharmacopoeia, the remedy is commonly used as an antitussive and expectorant. The shapes of young tubers from species of P. ternata are similar to those of P. pedatisecta and Arisaema heterophyllum, but different in medicinal properties. In order to provide molecular evidence for genuine origin identification of P. ternata species, the mannose-binding lectin sequences of P. ternata and its adulterants P. pedatisecta and A. heterophyllum were cloned using genomic walker technology. Based on the sequence analyses, we designed a pair of species-specific primers to authenticate P. ternata. For PCR-selective restriction (PCR-SR), we identified two distinctive sites which can be recognized by the restriction endonucleases BAMHI and NCOI in the open reading frame sequences of P. ternata, P. pedatisecta and A. heterophyllum. Our results indicate that the methods of PCR and PCR-SR are effective, accurate and applicable for identification of the bulbs of P. ternata.
    Planta Medica 08/2006; 72(9):844-7. · 2.35 Impact Factor
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    ABSTRACT: A full-length cDNA encoding a dehydrin was cloned from the living fossil plant Ginkgo biloba by rapid amplification of cDNA ends (RACE). The cDNA, designated as GbDHN, was 813 bp long containing an open reading frame of 489 bp. The deduced GbDHN protein had 163 amino acid residues, which formed a 17 kDa polypeptide with a predicted isoelectric point (pI) of 5.75. GbDHN had an S-segment and a K-segment, indicative of dehydrins, but no Y-segments. Homology analysis indicated that the S-segment and K-segment of GbDHN shared identity with those of other reported dehydrins, indicating that GbDHN belonged to dehydrin superfamily. Genomic sequence of GbDHN was also cloned using genomic walker technology. By comparing genomic DNA with the cDNA, it was found that there was a 257-bp intron in this gene. Promoter analysis indicated that it contained six CAAT boxes, one TATA box, one ABRE box and one GC-motif in the 5'-flanking region. Southern blot analysis revealed that GbDHN belonged to a single copy gene family. RT-PCR analysis revealed that GbDHN constitutively expressed in stems and roots. The increased expression of GbDHN was detected when G. biloba seedlings were treated with exogenous abscisic acid (ABA), salt stress and drought stress. These results indicate that the GbDHN has the potential to play a role in response to ABA and environmental stresses that can cause plant dehydration.
    Bioscience Reports 07/2006; 26(3):203-15. · 1.88 Impact Factor
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    ABSTRACT: A new gene, designated as BnPrx (GenBank Accession No. DQ078754), was isolated from oilseed rape (Brassica napus) by SMART Rapid Amplification of cDNA Ends (RACE). The full-length cDNA is 1307 bp long and contains a 1062 bp open reading frame (ORF), which encodes a 354 amino acid peroxidase precursor, with a 31 aa N-terminal signal peptide and a 15 aa C-terminal propeptide. The putative protein has a molecular weight of 38.86 kDa and a calculated pI of 5.85. BnPrx shares high identity with HRPC (89%). BnPrx possesses all active residues and two Ca(2+) sites present in Horseradish peroxidase isoenzymes C (HRPC) as well as six N-glycosylation sites. The predicted 3-D structure of BnPrx is very similar to that of HRPC. Assisted by genomic walking technology, the genomic DNA of BnPrx was also cloned, consisting of 3 introns and 4 exons. Thirty-two TATA boxes, 18 CAAT boxes and many cis-elements, such as WUN, MeJR, were found in its promoter region. Southern blot analysis indicated that BnPrx belonged to a small gene family. Northern blot analysis revealed that BnPrx was constitutively expressed in all tested tissues, including roots, stems and leaves, with the high expression in leaves and stems. The expression of BnPrx could be induced by methyl jasmonate (MeJA), salicylic acid (SA), cold and H(2)O(2). The cloning and characterizing of BnPrx might not only help us understand the physiological function and molecular evolution of the large peroxidase gene family more comprehensively, but also provide an alternative way of seeking a more effective and economical substitute for HRPC.
    Bioscience Reports 07/2006; 26(3):263-80. · 1.88 Impact Factor
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    ABSTRACT: 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR, EC: 1.1.1.34) catalyzes the first committed step in mevalonic acid (MVA) pathway for biosynthesis of isoprenoids. The full-length cDNA encoding HMGR was isolated from Ginkgo biloba for the first time (designated as GbHMGR, GenBank accession number AY741133), which contained a 1713 bp ORF encoding 571 amino acids. The GbHMGR genomic DNA sequence was also obtained, revealing GbHMGR had four exons and three introns. The deduced GbHMGR protein showed high identity to other plant HMGRs and contained two trans-membrane domains and a catalytic domain. The three dimensional model of GbHMGR represented a typical spatial structure of HMGRs. The Southern blot and RT-PCR assay results indicated that GbHMGR belonged to a small gene family, and expressed in a tissue-specific manner with a low level expression being only found in root. The potential significance of GbHMGR gene was also discussed.
    Molecular Biology Reports 07/2006; 33(2):117-27. · 2.51 Impact Factor
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    ABSTRACT: In many organisms, trehalose acts as protective metabolite against harsh environmental stresses, such as freezing, drought, nutrient starvation, heat and salt. Herein a cDNA (designated as GbTPS, GenBank Accession Number AY884150) encoding a trehalose-6-phosphate synthase homologue was isolated and characterized from the living fossil plant, Ginkgo biloba, which is highly tolerant to drought and cold. GbTPS encoded an 868-amino-acid polypeptide with a predicted isoelectric point of 5.83 and molecular mass of 97.9 kD. Amino acid sequence alignment revealed that GbTPS shared high identity with class II trehalose-6-phosphate synthase homologues (67% identical to AtTPS7), but had only 17% and 23% of identity with OstA from Escherichia coli and ScTPS1 from S. cerevisiae, respectively. DNA gel blot analysis indicated that GbTPS belonged to a small multi-gene family. The expression analysis by RT-PCR showed that GbTPS expressed in a tissue-specific manner in G. biloba and might involve in leaf development. GbTPS was also found to be induced by a variety of stresses including cold, salt, drought and mannitol.
    Journal of biochemistry and molecular biology 04/2006; 39(2):158-66. · 2.02 Impact Factor
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    ABSTRACT: Mevalonate diphosphate decarboxylase (MVD; EC4.1.1.33) catalyses the conversion of mevalonate diphosphate to isopentenyl diphosphate (IPP), which is a key skeleton for numerous functionally important secondary metabolites. A full-length cDNA of MVD was isolated from Ginkgo biloba (designated as GbMVD) using rapid amplification of cDNA ends (RACE). GbMVD was 1958 bp with poly(A) tailing and contained a 1290-bp open reading frame encoding a 430-amino acid protein. The deduced GbMVD protein had a deduced molecular weight of about 48 kDa and isoelectric point of 5.83, and it showed high similarity to MVDs from other species. The 3D-structure modelling showed that the 3D model of GbMVD had a similar P loop as that of yeast. Phylogenetic tree analysis revealed that GbMVD shared the same ancestor with MVDs from other species, and it had a closer relationship with those from plant species. Southern blot analysis indicated that GbMVD might belong to a small multigene family. Transcript accumulation analysis revealed that GbMVD was transcribed in root, stem and leaf tissues. Functional complementation of GbMVD in an MVD-deficient Saccharomyces cerevisiae strain MN19-34 (erg19ts) confirmed that GbMVD mediated the conversion of mevalonate diphosphate to IPP and that it is a functional gene.
    Physiologia Plantarum 03/2006; 127(1):19 - 27. · 3.66 Impact Factor
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    ABSTRACT: Flavanone 3-hydroxylase (F3H) activity is necessary for the biosynthesis of flavonoids, the main ingredients of Gingko biloba extract. The full-length cDNA and genomic DNA sequences of F3H gene were isolated from G. biloba for the first time. The full-length cDNA of G. biloba F3H gene (designated as GbF3H) contained a 1071 bp open reading frame (ORF) encoding a 357-amino-acid protein with a calculated molecular weight of about 40 kDa and isoelectric point (pI) of 5.57. The genomic DNA analysis showed that GbF3H gene had three exons and two introns. The deduced GbF3H protein showed high identities to other plant F3Hs. The conserved amino acids ligating ferrous iron and residues participating in 2-oxoglutarate binding (R-X-S) were found in GbF3H at the similar positions like other F3Hs. Three-dimensional structure modeling showed that GbF3H had a jerry roll in the enzyme core consisted of beta-sheet, a typical structure shared by all 2-oxoglutarate-dependent dioxygenases including F3Hs. Phylogenetic tree analysis revealed that GbF3H shared the same ancestor in evolution with other F3Hs and had a further relationship with other angiosperms species. Southern blot analysis indicated that GbF3H belonged to a multi-gene family. Transcription analysis revealed that GbF3H expressed in stem and leaf with the highest transcription level in leaf. The isolation and characterization of GbF3H gene will be helpful to further study the role of GbF3H gene in the biosynthesis of flavonoids in G. biloba.
    Bioscience Reports 03/2006; 26(1):19-29. · 1.88 Impact Factor
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    ABSTRACT: We isolated a putative anthocyanidin reductase gene from Ginkgo biloba (designated as GbANR) by rapid amplification of cDNA ends (RACE). The full-length cDNA of GbANR is 1451 bp long with poly(A) tail and contains a 1026 bp open reading frame (ORF) encoding 342 amino acids. The GbANR gene is composed of five exons and four introns. Sequence alignment shows that the deduced GbANR has a relatively high identity to other plant anthocyanidin reductases at the amino acid level. DNA gel blot analysis indicates that GbANR belongs to a small gene family. Transcription analysis indicates that GbANR is preferentially expressed in leaves.
    Journal of Plant Physiology 03/2006; 163(2):224-7. · 2.70 Impact Factor
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    ABSTRACT: A novel defensin gene was isolated from Ginkgo biloba. The full-length cDNA of G. biloba defensin (designated as Gbd) was 534bp. The cDNA contained a 240-bp open reading frame encoding an 80-amino acid protein of 5.68 kDa with a potential 30 aa signal peptide. The putative GbD mature protein showed striking similarity to other plant defensins, representing low molecular size antimicrobial polypeptides. Eight cysteine sites conserved in plant defensins were also found in GbD at similar positions. Three-dimensional structure modeling showed that GbD strongly resembled defensin from tobacco (NaD1) and consisted of an alpha-helix and a triple-strand antiparallel beta-sheet that were stabilized by four intramolecular disulfide bonds, implying GbD may have functions similar to NaD1. The genomic DNA gel blot indicated that Gbd belonged to a multigene family. Expression analysis revealed that Gbd was up-regulated by wounding and methyl jasmonate treatments, suggesting that Gbd is potentially involved in plant resistance or tolerance to pathogens during wounding.
    Journal of Plant Physiology 11/2005; 162(10):1160-8. · 2.70 Impact Factor
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    ABSTRACT: A new abscisic acid, stress and ripening (Asr) gene was cloned from Ginkgo biloba by rapid amplification of cDNA ends (RACE) method. The full-length cDNA of G. biloba Asr (designated as GbAsr) was 952 bp long and it contained a 543 bp open reading frame encoding a protein of 181 amino acids. GbASR was found to be rich in His, Lys, Glu and Ala, and it had extensive homology with those of other plant Asr genes via multiple alignment analysis. Phylogenetic tree analysis indicated that the GbASR had a closer relationship with ASR from pine, another gymnosperm species, than with angiosperm ASRs. Southern blot analysis indicated that GbAsr belonged to a small multigene family. RT-PCR analyses revealed that GbAsr had a distinct up-regulated transcript pattern in root, stem and leaf under mannitol, NaCl and ABA treatments. The recombinant GbASR protein was successfully expressed in E. coli strain with pET-32a vector, and the result showed that the molecular weight of the recombinant protein was about 20 kDa, a size in agreement with that of the predicted by bioinformatic analysis. The expression of the GbAsr in E. coli will facilitate further research on this gene.
    Plant Physiology and Biochemistry 10/2005; 43(9):836-43. · 2.78 Impact Factor
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    ABSTRACT: The wilt defense reaction of cotton is a complicated continuous process and involves a battery of genes. In this study, we adopted the suppression subtractive hybridization (SSH) technique to isolate differentially expressed ESTs from Gossypium barbadense variety 7124 during the Verticillium wilt defense process. An array of 1165 clones from the subtractive library has been screened with reverse northern blotting, of which 131 ESTs were considered as overexpressed and 16 ESTs were downregulated. Sequence analysis and blast search showed that 83 ESTs were homologous to 45 unique sequences in the databases. Among all these differentially expressed ESTs, at least three kinds of genes were characterized. The majority of ESTs with a deduced identity as aerobic metabolism enzymes were strongly expressed in the infection process. Likewise, ESTs similar to those reported for pathogen-related protein genes were also picked out in this study. These ESTs, in combination with other kinase-like genes and a defensin-like EST, constituted an assembly of genes which responded during pathogenic infection. These results imply that sea-island cotton undergoes strong oxidative stress and results in a series of defense responses when attacked by V. dahliae. To our knowledge, this is the first report on the isolation of global ESTs during the sea-island cotton defense reaction.
    Molecular Biology 02/2005; 39(2):191-199. · 0.64 Impact Factor
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    ABSTRACT: The genomic DNA sequence of chalcone synthase (CHS) gene was cloned from Ginkgo biloba. The Gbchs was 1295 bp long and composed of two exons and one intron, one of the typical features of chalcone synthase genes. The genomic Southern blot analysis indicated that Gbchs belonged to a multigene family. RT-PCR analyses revealed that Gbchs expressed differentially in the root, stem and leaf tissues of G. biloba, and the expression was induced by UV-B and wounding treatments. The recombinant GbCHS protein was successfully expressed in Escherichia coli strain M15 [pREP4] with pQE30 vector and the result showed that the expressed GbCHS protein had molecular weight of about 42 kDa, a size matching that of the predicted by bioinformatic analysis. This study provides useful information for further studying this gene and its function in ginkgo flavonoids biosynthetic pathway in G. biloba, the so-called “living fossil” plant possessing many pharmaceutical properties for human health.
    Plant Science. 01/2005;
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    ABSTRACT: A full-length cDNA of a new serine/threonine (Ser/Thr) protein kinase gene, designated as BnSOS2 (GenBank Acc. No.AY310413), was cloned from Brassica napus by rapid amplification of cDNA ends (RACE). The full-length cDNA of BnSOS2 was 1779 bp and contained a 1539-bp open reading frame encoding a protein of 512 amino acids. Homology analysis shows that BnSOS2 strongly resembles other Ser/Thr protein kinase genes, and that its putative protein belongs to a typical Ser/Thr kinase family. Northern blot analysis reveals that BnSOS2 is salt-inducible. Our results indicate that BnSOS2 is a new member of the plant SOS2 gene family, which may play an important role in salt tolerance of plants.
    Cellular & Molecular Biology Letters 02/2004; 9(3):465-73. · 1.95 Impact Factor
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    ABSTRACT: A new Na+/H+ vacuolar antiporter gene from Brassica napus was cloned. The full-length cDNA of B. napus antiporter gene (BnNHX1, GenBank Acc. No. AY189676) was 1819 bp and contained a 1545 bp open reading frame encoding a protein of 515 amino acids. Homology analysis and molecular modeling revealed that BnNHX1 strongly resembled other Na+/H+ antiporter genes. Its encoded protein belonged to a typical sodium/hydrogen exchanger superfamily and shared consensus sequences within predicted transmemberane segments. Southern blot analysis indicated there were multi-copies of BnNHX1 gene present in B. napus genome, which was different from that in Oryza sativa previously reported. Northern blot analysis revealed that BnNHX1 was salt-inducible and its transcript level was most abundant after 24 h treatment with 200 mM sodium chloride. Our studies suggested that BnNHX1 was a new member of the family of recently cloned plant NHX-genes.
    DNA Sequence 11/2003; 14(5):351-8. · 0.75 Impact Factor

Publication Stats

129 Citations
31.94 Total Impact Points

Institutions

  • 2007
    • Shanghai Academy of Public Measurement
      Shanghai, Shanghai Shi, China
  • 2003–2007
    • Fudan University
      • • State Key Laboratory of Genetic Engineering
      • • School of Life Sciences
      Shanghai, Shanghai Shi, China
  • 2005–2006
    • Shanghai Jiao Tong University
      • School of Agriculture and Biology
      Shanghai, Shanghai Shi, China