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1.
cDNA cloning and characterization of a mannose-binding lectin fromZingiber officinaleRoscoe (ginger) rhizomes 总被引:1,自引:0,他引:1
Using RNA extracted fromZingiber officinale rhizomes and primers designed according to the conservative regions of monocot mannose-binding lectins, the full-length cDNA
ofZ. officinale agglutinin (ZOA) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA ofzoa was 746 bp and contained a 510 bp open reading frame (ORF) encoding a lectin precursor of 169 amino acids with a signal peptide.
ZOA was a mannose-binding lectin with three typical mannose-binding sites (QDNY). Semi-quantitative RT-PCR analysis revealed
thatzoa expressed in all the tested tissues ofZ. officinale including leaf, root and rhizome, suggesting it to be a constitutively expressing form. ZOA protein was successfully expressed
inEscherichia coli with the molecular weight expected. To our knowledge, this is the first mannose-binding lectin cDNA cloned from the family
Zingiberaceae. Our results demonstrate that monocot mannose-binding lectins also occur within the family Zingiberaceae 相似文献
2.
Using RNA extracted from Pinellia cordata young leaves and primers designed according to the conserved regions of Araceae lectins, the full-length cDNA of Pinellia cordata agglutinin (PCL) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of pcl was 1,182 bp and contained a 768 bp open reading frame (ORF) encoding a lectin precursor of 256 amino acids. Through comparative
analysis of pcl gene and its deduced amino acid sequence with those of other Araceae species, it was found that pcl encoded a precursor lectin with signal peptide. PCL is a mannose-binding lectin with three mannose-binding sites. Semi-quantitative
RT-PCR analysis revealed that pcl is expressed in all tested tissues including leaf, stem and bulbil, but with the highest expression in bulbil. PCL protein
was successfully expressed in Escherichia coli with the molecular weight expected. 相似文献
3.
A new lectin gene was isolated by using genomic walker technology and revealed to encode a mannose-binding lectin. Analysis of a 2233 bp segment revealed a gene including a 1169 bp 5′ flanking region, a 417 bp open reading frame (ORF) and a 649 bp 3′ flanking region. There are two putative TATA boxes and eight possible CAAT boxes lie in the 5′ flanking region. The ORF encodes a 15.1 kDa precursor, which contains a 24-amino acid signal peptide. One possible polyadenylation signal is found in the 3′-flanking region. No intron was detected within the region of genomic sequence corresponding to zaa (Zantedeschia aethiopica agglutinin) full-length cDNA, which is typical of other mannose-binding lectin gene that have been reported. The deduced amino acid sequence of the lectin gene coding region shares 49–54% homology with other known lectins. The cloning of this new lectin gene will allow us to further study its structure, expression and regulation mechanisms. 相似文献
4.
cDNA cloning and expression analysis of a mannose-binding lectin from <Emphasis Type="Italic">Pinellia pedatisecta</Emphasis> 总被引:2,自引:0,他引:2
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-DOM1 (polypeptides A) and PPA-DOM2 (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. 相似文献
5.
A new lectin gene was cloned from Amorphophallus konjac. The full-length cDNA of Amorphophallus konjac agglutinin (aka) was 736 bp and contained a 474 bp open reading frame encoding a 158 amino acid protein. Homology analysis revealed that the lectin from this Araceae species belonged to the superfamily of monocot mannose-binding proteins. Molecular modeling of AKA indicated that the three-dimensional structure of AKA strongly resembles that of the snowdrop lectin. Southern blot analysis of the genomic DNA revealed that aka belonged to a low-copy gene family. Northern blot analysis demonstrated that aka expression was tissue-specific with the strongest expression being found in root. 相似文献
6.
Molecular cloning of the lectin and a lectin-related protein from common Solomon's seal (Polygonatum multiflorum) 总被引:3,自引:0,他引:3
Els J. M. Van Damme Annick Barre Pierre Rougé Fred Van Leuven Jan Balzarini Willy J. Peumans 《Plant molecular biology》1996,31(3):657-672
The most prominent protein ofPolygonatum multiflorum (common Solomon's seal) rhizomes has been identified as a mannose-binding lectin. Analysis of the purified lectin demonstrated that it is a tetramer of four identical subunits of 14 kDa. Molecular cloning further revealed that the lectin from this typical Liliaceae species belongs to the superfamily of monocot mannose-binding proteins. Screening of cDNA libraries constructed with RNA isolated from buds, leaves and flowers ofP. multiflorum also yielded cDNA clones encoding a protein, which contains two tandemly arranged domains with an obvious sequence homology to the mannose-binding lectins. Molecular modelling of thePolygonatum lectin and lectin-related protein indicated that the three-dimensional structure of both proteins strongly resembles that of the snowdrop lectin. In addition, this approach suggested that the presumed carbohydrate-binding sites of the lectin can accommodate a mannose residue whereas most of the carbohydratebinding sites of the lectin-related protein cannot.Abbreviations GNA
Galanthus nivalis agglutinin
- HCA
hydrophobic cluster analysis
- LECPMA
cDNA clone encoding PMA
- PM30
30 kDa protein isolated fromPolygonatum multiforum
- PMA
Polygonatum multiflorum agglutinin
- PMLRP
Polygonatum multiflorum lectin-related protein 相似文献
7.
Lin J Zhou X Pang Y Gao H Fei J Shen GA Wang J Li X Sun X Tang K 《Bioscience reports》2005,25(5-6):345-362
A novel agglutinin gene was cloned from Arisaema lobatum using SMART RACE-PCR technology. The full-length cDNA of Arisaema lobatum agglutinin (ala) was 1078 bp and contained a 774 bp open reading frame encoding a lectin precursor (proproprotein) of 258 amino acid residues
with a 23 aa signal peptide. ALA contained three mannose-binding sites (QXDXNXVXY) with two-conserved domains of 45% identity,
ALA-DOM1 and ALA-DOM2. The three-dimensional structure of ALA was very similar to that of GNA (Galanthus nivalis agglutinin). ALA shared varying identities, ranging from 40% to 85%, with mannose-binding lectins from other species of plant
families, such as Araceae, Alliaceae, Iridaceae, Lillaceae, Amaryllidaceae and Bromeliaceae. Genomic sequence of ala was also cloned using genomic walker technology, and it was found to contain three putative TATA boxes and eight possible
CAAT boxes in the 5′-flanking region. No intron was found within the region of genomic sequence. Southern blot analysis indicated
that the ala belonged to a multi-copy gene family. Expression pattern analysis revealed that the ala preferentially expressed in the tissues with the higher expression being found in spadix, bud, leaf, spathe and tuber. The
cloning of the ala gene not only provides a basis for further investigation of its structure, expression and regulation 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. 相似文献
8.
In this paper we report on the molecular cloning, sequencing and partially characterisation of a lectin from bulb of the Chinese
medicinal plant Zephyranthes grandiflora. The full-length cDNA of Z. grandiflora bulb lectin (ZGBL) consisted of 986 bp and contained a 576 bp ORF encoding a 191 amino acid protein. Bioinformatics analysis
results clearly indicate that ZGBL belongs to the monocot mannose-binding lectin family, which contains 3 putative mannose-binding
sites per subunit. RT-PCR analysis results indicate that ZGBL is constitutively expressed in all the tested tissue types including root, bulb, leaf and flower. Interestingly, ZGBL is
more closely related to the Orchidaceae rather than the Amaryllidaceae family on molecular evolution. 相似文献
9.
Zhang W Peumans WJ Barre A Astoul CH Rovira P Rougé P Proost P Truffa-Bachi P Jalali AA Van Damme EJ 《Planta》2000,210(6):970-978
A novel plant lectin was isolated from salt-stressed rice (Oryzasativa L.) plants and partially characterized. The lectin occurs as a natural mixture of two closely related isoforms consisting
of two identical non-covalently linked subunits of 15 kDa. Both isoforms are best inhibited by mannose and exhibit potent
mitogenic activity towards T-lymphocytes. Biochemical analyses and sequence comparisons further revealed that the rice lectins
belong to the subgroup of mannose-binding jacalin-related lectins. In addition, it could be demonstrated that the lectins
described here correspond to the protein products of previously described salt-stress-induced genes. Our results not only
identify the rice lectin as a stress protein but also highlight the possible importance of protein-carbohydrate interactions
in stress responses in plants.
Received: 27 July 1999 / Accepted: 11 November 1999 相似文献
10.
Shigeyuki Tsutsui Masaki Okamoto Satoshi Tasumi Hiroaki Suetake Kiyoshi Kikuchi Yuzuru Suzuki 《Comparative biochemistry and physiology. Part D, Genomics & proteomics》2006,1(1):122
In earlier work, we identified a novel mannose-specific lectin, termed pufflectin-s, from the skin mucus of torafugu, Takifugu rubripes. We here make a brief review on the lectin. The amino acid sequence of pufflectin-s shares sequence homology with mannose-binding lectins of monocotyledonous plants, and has conserved two of three carbohydrate recognition domains, QDNVY motifs, of these plant lectins. By site-directed mutagenesis, we verified that the QDNVY motif in the N-terminal region was critical to the mannose-binding function of pufflectin-s. RT-PCR and Northern blot analyses indicated that the pufflectin-s gene is expressed in the gill, oral cavity wall, esophagus, and skin. In addition, an isoform, pufflectin-i, which shares 91.4% amino acid identity with pufflectin-s, was isolated from the intestine. Using immunohistochemistry, pufflectin-s could be detected exclusively in the epithelial cells of the skin, gill, oral cavity wall and esophagus, whereas pufflectin-i was observed in both mucous and epithelial cells in the intestine. Nevertheless, mRNAs for both pufflectins were detected only in epithelial cells of these tissues with in situ hybridization. Pufflectin-s agglutinated some bacteria isolated from rearing water and from fish skin. This lectin also bound to a parasite, Heterobothrium okamotoi, suggesting that it may play an important role in the self-defense system of fugu. 相似文献
11.
To identify the genes involved in flower development, we analyzed 207 expressed sequence tags (ESTs) from a young floral bud
cDNA library ofPharbitis nil. Of these, 87 clones (42%) showed significant homology to known protein sequences in the NCBI database. Four of these had
not been reported previously in the plant kingdom, indicating that 1.9% of the ESTs were newly identified in plants. Functional
categorization revealed that the genes involved in metabolic pathways, such as glycolysis and photosynthesis, were most abundant
Reverse-northern and northern analyses showed that one clone,PnFP161, was expressed preferentially in floral buds. DNA sequence analysis indicated that this clone contained 147 bp of 5′-UTR,
264 bp of -UTR, and an open reading frame of 233 amino acids, thereby sharing 33% identity with a lectin fromCalystegia sepium. The C-terminal regionof PnFPI61 had well-conserved residue with that of the lectins. Southern blot analysis demonstrated thatPnFPI61 exists as a multigene family. 相似文献
12.
Summary Mannose/glucose- and galactose-binding lectins (ML and GL respectively, were located by immunogold labelling in tissues of a peanut (Arachis hypogaea) nodule induced by an effectiveBradyrhizobium sp. strain. Light and electron microscopic examination of silver-enhanced semithin and ultrathin sections, respectively, revealed that both lectins were widely distributed throughout the cortex and bacteroidal zones although ML was more abundant. The lectins were predominantly in the vacuoles of cortical cells but GL was absent from, or at low concentration in, a two-cell-thick layer of cortical cells surrounding the bacteroidal region. Only ML was detected in cells of the vascular bundle endodermis and in central vascular bundle cells; neither lectin was found in pericycle cells. Bacteroidal cells contained abundant ML in the nuclei and cytoplasm surrounding bacteroids while GL was mainly located in the central vacuoles of these cells. Neither lectin was associated with bacteroid surfaces, peribacteroid membranes, plant cell walls or cell organelles and membranes. The above observations indicate that the nodule lectins are not symbiotic cell recognition determinants and suggest that they have protein storage functions.Abbreviations BSA
bovine serum albumin
- GL
galactose-binding lectin
- ML
mannose-binding lectin
- PBS
phosphate-buffered saline
- PBST
phosphate-buffered saline plus Tween 相似文献
13.
Sadeghi A Smagghe G Broeders S Hernalsteens JP De Greve H Peumans WJ Van Damme EJ 《Transgenic research》2008,17(1):9-18
The insecticidal activity of the leaf (ASAL) and bulb (ASAII) agglutinins from Allium sativum L. (garlic) against the cotton leafworm, Spodoptera littoralis Boisd. (Lepidoptera: Noctuidae) was studied using transgenic tobacco plants expressing the lectins under the control of the
constitutive CaMV35S promoter. PCR analysis confirmed that the garlic lectin genes were integrated into the plant genome.
Western blots and semi-quantitative agglutination assays revealed lectin expression at various levels in the transgenic lines.
Biochemical analyses indicated that the recombinant ASAL and ASAII are indistinguishable from the native garlic lectins. Insect
bioassays using detached leaves from transgenic tobacco plants demonstrated that the ectopically expressed ASAL and ASAII
significantly (P < 0.05) reduced the weight gain of 4th instar larvae of S. littoralis. Further on, the lectins retarded the development of the larvae and their metamorphosis, and were detrimental to the pupal
stage resulting in weight reduction and lethal abnormalities. Total mortality was scored with ASAL compared to 60% mortality
with ASAII. These findings suggest that garlic lectins are suitable candidate insect resistance proteins for the control of
S. littoralis through a transgenic approach. 相似文献
14.
Suzuki Y Tasumi S Tsutsui S Okamoto M Suetake H 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2003,136(4):723-730
Among lectins in the skin mucus of fish, primary structures of four different types of lectin have been determined. Congerin from the conger eel Conger myriaster and AJL-1 from the Japanese eel Anguilla japonica were identified as galectin, characterized by its specific binding to β-galactoside. Eel has additionally a unique lectin, AJL-2, which has a highly conserved sequence of C-type lectins but displays Ca2+-independent activity. This is rational because the lectin exerts its function on the cutaneous surface, which is exposed to a Ca2+ scarce environment when the eel is in fresh water. The third type lectin is pufflectin, a mannose specific lectin in the skin mucus of pufferfish Takifugu rubripes. This lectin showed no sequence similarity with any known animal lectins but, surprisingly, shares sequence homology with mannose-binding lectins of monocotyledonous plants. The fourth lectin was found in the ponyfish Leiognathus nuchalis and exhibits homology with rhamnose-binding lectins known in eggs of some fish species. These lectins, except ponyfish lectin, showed agglutination of certain bacteria. In addition, pufflectin was found to bind to a parasitic trematode, Heterobothrium okamotoi. Taken together, these results demonstrate that skin mucus lectins in fish have wide molecular diversity. 相似文献
15.
Ponpimol Tipthara Polkit Sangvanich Marcus Macth Amorn Petsom 《Journal of Plant Biology》2007,50(2):167-173
A mannose-binding lectin was isolated from rhizomes of the medicinal plantCurcuma zedoaria. We used extraction with 20 mM phosphate buffer, ammonium sulfate precipitation, ion exchange chromatography on Q-Sepharose,
gel filtration chromatography on Superdex 75, and reverse-phase HPLC. The purified lectin yielded a single band on SDS-PAGE
that corresponded to a molecular mass of 13 kDa. This lectin exhibited hemagglutinating activity toward rabbit erythrocytes,
which could be inhibited by mannose only. The lectin was digested with trypsin and its digests were analyzed using MALDI-TOF/TOF.
Partial amino acid sequences were obtained from tandem mass spectra via automatedde novo sequencing, and were then identified by MS-BLAST homology searches to enable recognition of related proteins in other species.
Inferred peptide sequences exhibited similarity to a mannose-binding lectin fromEpipactis helleborine, a member of the Orchidaceae. 相似文献
16.
Els J. M. van Damme Jan Balzarini Koen Smeets Fred van Leuven Willy J. Peumans 《Glycoconjugate journal》1994,11(4):321-332
The Orchidaceae speciesListera ovata andEpipactis helleborine contain two types of mannose-binding proteins. Using a combination of affinity chromatography on mannose-Sepharose-4B and ion exchange chromatography on a Mono-S column eight different mannose-binding proteins were isolated from the leaves ofListera ovata. Whereas seven of these mannose-binding proteins have agglutination activity and occur as dimers composed of lectin subunits of 11–13 kDa, the eighth mannose-binding protein is a monomer of 14 kDa devoid of agglutination activity. Moreover, the monomeric mannose-binding protein does not react with an antiserum raised against the dimeric lectin and, in contrast to the lectins, is completely inactive when tested for antiretroviral activity against human immunodeficiency virus type 1 and type 2. Mannose-binding proteins with similar properties were also found in the leaves ofEpipactis helleborine. However, in contrast toListera only one lectin was found inEpipactis. Despite the obvious differences in molecular structure and biological activities molecular cloning of different mannose-binding proteins fromListera andEpipactis has shown that these proteins are related and some parts of the sequences show a high degree of sequence homology indicating that they have been conserved through evolution.Abbreviations EHMBP
Epipactis helleborine mannose-binding protein
- LOMBP
Listera ovata mannose-binding protein
Note: The nucleotide sequences reported in this paper will appear in the Genbank/EMBL Data library with the accession numbers L18894, L18895 and U07787. 相似文献
17.
18.
Zhonghai Chen Xiaofen Sune Kexuan Tange 《Journal of plant biochemistry and biotechnology.》2005,14(1):33-36
Using RNA extracted from Dendrobium officinale young leaves and primers designed according to the conservative regions of Orchidaceae lectins, the full-length cDNA of Dendrobium officinale agglutinin (DOA) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of doa was 768 bp and contained a 498 bp open reading frame (ORF) encoding a lectin precursor of 165 amino acids. Through comparative analysis of doa gene and its deduced amino acid sequence with those of other Orchidaceae species, it was found that doa encoded a precursor lectin with signal peptide. DOA was a mannose-binding lectin with three mannose-binding sites. Semi-quantitative RT-PCR analysis revealed that doa mRNA expression was detected in all tested tissues including root, stem and leaf, however, the expression was higher in stem, lower in leaf. As the doa mRNA was detected in all the tested plant tissues, the doa was considered to be a constitutively expressed gene. 相似文献
19.
Els J. M. Van Damme Koen Smeets Iris Engelborghs Helen Aelbers Jan Balzarini Arpad Pusztai Fred van Leuven Irwin J. Goldstein Willy J. Peumans 《Plant molecular biology》1993,23(2):365-376
Characterization of the lectins from onion (Allium cepa), shallot (A. ascalonicum) and leek (A. porrum) has shown that these lectins differ from previously isolated Alliaceae lectins not only in their molecular structure but also in their ability to inhibit retrovirus infection of target cells.cDNA libraries constructed from poly(A)-rich RNA isolated from young shoots of onion, shallot and leek were screened for lectin cDNA clones using colony hybridization. Sequence analysis of the lectin cDNA clones from these three species revealed a high degree of sequence similarity both at the nucleotide and at the amino acid level.Apparently the onion, shallot and leek lectins are translated from mRNAs of ca. 800 nucleotides. The primary translation products are preproproteins (ca. 19 kDa) which are converted into the mature lectin polypeptides (12.5–13 kDa) after post-translational modifications.Southern blot analysis of genomic DNA has shown that the lectins are most probably encoded by a family of closely related genes which is in good agreement with the sequence heterogeneity found between different lectin cDNA clones of one species. 相似文献
20.
We have characterized pufflectin, a novel mannose-specific lectin, from the skin mucus of the pufferfish, Fugu rubripes. Molecular mass estimations by gel filtration and matrix-assisted laser desorption ionization time-of-flight mass spectrometry and the SDS-PAGE pattern suggest that pufflectin is a homodimer composed of non-covalently associated subunits of 13 kDa. The full-length pufflectin cDNA consists of 527 bp, with 116 amino acid residues deduced from the open reading frame. The amino acid sequence of pufflectin shows no homology with any known animal lectin. Surprisingly, pufflectin shares sequence homology with mannose-binding lectins of monocotyledonous plants and has conserved two of three carbohydrate recognition domains of these plant lectins. The pufflectin gene is expressed in gills, oral cavity wall, esophagus, and skin. In addition, an isoform occurs exclusively in the intestine. Pufflectin differs from mannose-binding lectins purified from the blood plasma of Fugu. Whereas pufflectin did not agglutinate five bacterial species tested, it was demonstrated to bind to the parasitic trematode, Heterobothrium okamotoi. This finding suggests that pufflectin contributes to the parasite-defense system in Fugu. 相似文献