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  • 1
    Electronic Resource
    Electronic Resource
    Constitutive nitrogenase synthesis from de novo transcribed mRNA in isolated Rhizobium leguminosarum bacteroids
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Gene Structure and Expression 740 (1983), S. 313-322 
    Details
    ISSN: 0167-4781
    Keywords: (R. leguminosarum) ; Bacteroid ; Gene expression ; Nitrogenase synthesis ; Translational regulation ; mRNA half-life
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
    URL: http://linkinghub.elsevier.com/retrieve/pii/0167-4781(83)90140-9
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  • 2
    Electronic Resource
    Electronic Resource
    Plant degradation A nematode expansin acting on plants (2004)
    [s.l.] : Nature Publishing Group
    Nature 427 (2004), S. 30-30 
    Details
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Expansin proteins, which have so far been identified only in plants, rapidly induce extension of plant cell walls by weakening the non-covalent interactions that help to maintain their integrity. Here we show that an animal, the plant-parasitic roundworm Globodera rostochiensis, can also ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/427030a
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  • 3
    Electronic Resource
    Electronic Resource
    Enzymology Degradation of plant cell walls by a nematode (2000)
    [s.l.] : Macmillan Magazines Ltd.
    Nature 406 (2000), S. 36-37 
    Details
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Interwoven networks of cellulose and pectin are the main components of plant cell walls, making them recalcitrant structures that can only be degraded by organisms producing a mix of synergistically acting enzymes. Animals were believed to be unable to synthesize these enzymes, depending ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/35017641
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  • 4
    Electronic Resource
    Electronic Resource
    NEMATODE PARASITISM GENES (2000)
    Palo Alto, Calif. : Annual Reviews
    Annual Review of Phytopathology 38 (2000), S. 365-396 
    Details
    ISSN: 0066-4286
    Source: Annual Reviews Electronic Back Volume Collection 1932-2001ff
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Biology
    Notes: Abstract The ability of nematodes to live on plant hosts involves multiple parasitism genes. The most pronounced morphological adaptations of nematodes for plant parasitism include a hollow, protrusible stylet (feeding spear) connected to three enlarged esophageal gland cells that express products that are secreted into plant tissues through the stylet. Reverse genetic and expressed sequence tag (EST) approaches are being used to discover the parasitism genes expressed in nematode esophageal gland cells. Some genes cloned from root-knot (Meloidogyne spp.) and cyst (Heterodera and Globodera spp.) nematodes have homologues reported in genomic analyses of Caenorhabditis elegans and animal-parasitic nematodes. To date, however, the candidate parasitism genes endogenous to the esophageal glands of plant nematodes (such as the ss-1,4-endoglucanases) have their greatest similarity to microbial genes, prompting speculation that genes for plant parasitism by nematodes may have been acquired by horizontal gene transfer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1146/annurev.phyto.38.1.365
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  • 5
    Electronic Resource
    Electronic Resource
    Coordinate expression of antibody subunit genes yields high levels of functional antibodies in roots of transgenic tobacco (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1701-1710 
    Details
    ISSN: 1573-5028
    Keywords: Antibody ; coordinated expression ; genetic engineering ; protein assembly ; root ; secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To explore the feasibility of employing antibodies to obtain disease resistance against plant root pathogens, we have studied the expression of genes encoding antibodies in roots of transgenic plants. A model monoclonal antibody was used that binds to a fungal cutinase. Heavy and light chain cDNAs were amplified by PCR, fused to a signal sequence for secretion and cloned behind CaMV 35S and TR2′ promoters in a single T-DNA. The chimeric genes were cloned both in tandem and in a divergent orientation. The roots of tobacco plants transformed with these constructs produced antibodies that were able to bind antigen in an ELISA. Immunoblotting showed assembly to a full-size antibody. In addition, a F(ab′)2-like fragment was observed, which is probably formed by proteolytic processing. Both antibody species were properly targeted to the apoplast, but the full-size antibody was partially retained by the wall of suspension cells. The construct with divergent promoters showed a better performance than the construct with promoters in tandem. It directed the accumulation of functional antibodies to a maximum of 1.1% of total soluble protein, with half of the plants having levels higher than 0.35%. The high efficiency of this construct probably results from coordinated and balanced expression of light and heavy chain genes, as evidenced by RNA blot hybridization.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019485
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  • 6
    Electronic Resource
    Electronic Resource
    The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco (1996)
    Springer
    Plant molecular biology 30 (1996), S. 781-793 
    Details
    ISSN: 1573-5028
    Keywords: Genetic engineering ; KDEL retention signal ; signal peptide ; single-chain antibody ; targeting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of subcellular localization on single-chain antibody (scFv) expression levels in transgenic tobacco was evaluated using an scFv construct of a model antibody possessing different targeting signals. For translocation into the secretory pathway a secretory signal sequence preceded the scFv gene (scFv-S). For cytosolic expression the scFv antibody gene lacked such a signal sequence (scFv-C). Also, both constructs were provided with the endoplasmic reticulum (ER) retention signal KDEL (scFv-SK and scFv-CK, respectively). The expression of the different scFv constructs in transgenic tobacco plants was controlled by a CaMV 35S promoter with double enhancer. The scFv-S and scFv-SK antibody genes reached expression levels of 0.01% and 1% of the total soluble protein, respectively. Surprisingly, scFv-CK transformants showed considerable expression of up to 0.2% whereas scFv-C transformants did not show any accumulation of the scFv antibody. The differences in protein expression levels could not be explained by the steady-state levels of the mRNAs. Transient expression assays with leaf protoplasts confirmed these expression levels observed in transgenic plants, although the expression level of the scFv-S construct was higher. Furthermore, these assays showed that both the secretory signal and the ER retention signal were recognized in the plant cells. The scFv-CK protein was located intracellularly, presumably in the cytosol. The increase in scFv protein stability in the presence of the KDEL retention signal is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019011
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  • 7
    Electronic Resource
    Electronic Resource
    Selection of Beet Necrotic Yellow Vein Virus Specific Single-chain Fv Antibodies from a Semi-synthetic Combinatorial Antibody Library (1999)
    Springer
    European journal of plant pathology 105 (1999), S. 147-156 
    Details
    ISSN: 1573-8469
    Keywords: BNYVV ; ELISA ; phage-antibodies ; phage display ; ScFv ; ScFv-AP/S ; sugar beet
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Methods for the generation of monoclonal antibodies against plant viruses are limited because current hybridoma techniques do not allow efficient exploitation of the immune repertoire. Moreover, the immunization procedures often lead to a bias towards an immunodominant contaminant in the immunogen preparation and not to the plant virus itself. The selection of six different single-chain antibody variable fragments (scFv) against beet necrotic yellow vein virus from a semi-synthetic human combinatorial antibody library showed the feasibility of the phage display system. No bias towards minor contaminants in the purified virus preparation was observed in ELISA, as all the selected scFvs reacted only with beet necrotic yellow vein virus infected plant homogenates. In addition, two of the isolated beet necrotic yellow vein virus-specific scFvs could be produced in E. coli as a scFv fusion protein with alkaline phosphatase, and were applied in ELISA as specific ready to use antibody-enzyme conjugates. Because of their specificity, these antibodies have potential to be used as reagents in sensitive diagnostic assays for routine testing for beet necrotic yellow vein virus in sugar beets.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008727326113
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  • 8
    Electronic Resource
    Electronic Resource
    Specificity of polycllonal and monoclonal antibodies for the identification of Xanthomonas campestris pv. campestris (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 81-94 
    Details
    ISSN: 1573-8469
    Keywords: black rot ; serology ; sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) ; proteinase K
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Polyclonal and monoclonal antibodies (PCAs and MCAs), produced to whole cells and flagellar extracts ofXanthomonas campestris pv.campestris (Xcc), respectively, were tested for specificity. In immunofluorescence microscopy (IF) the three PCAs tested, reacted at low dilutions with all Xcc strains, some other xanthomonads and non-xanthomonads. At higher dilutions most cross-reactivity with non-xanthomonad strains disappeared. However, the cross-reactivity with strains ofX. c. pv.vesicatoria (Xcv),X. c. pv.amoraciae (Xca) andX. c. pv.phaseoli var. fuscans (Xcpf) remained. Six MCA-producing cell clones viz. 20H6, 2F4, 18G12, 10C5, 17C12 and 16B5 were selected for specificity tests with an enzyme immunoassay (EIA), IF and a dot-blot immunoassay (DBI). None of the MCAs reacted with all Xcc strains in IF and EIA. In DBI, only MCAs 17C12 and 16B5 reacted with all Xcc strains. All six MCAs tested, cross-reacted in one of either tests with other pathovars ofX. campestris, such as Xcv or Xca. The MCAs were also tested in immunoblotting experiments using total bacterial extracts, cell envelope and flagellar extracts. MCAs 20H6, 2F4, 18G12 and 10C5 reacted with the lipopolysaccharide (LPS) of Xcc. MCAs 16B5 and 17C12 reacted with a 39 kilodalton and a 29 kilodalton protein, respectively. It is concluded that the PCAs and MCAs discussed in this study may be used for routine identification and differentiation of (a group of) Xcc strains. The significance of the cross-reactions with other pathovars ofX. campestris needs to be determined by testing seed lots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01996321
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  • 9
    Electronic Resource
    Electronic Resource
    ‘Plantibodies’: a flexible approach to design resistance against pathogens (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 183-191 
    Details
    ISSN: 1573-8469
    Keywords: Monoclonal antibodies ; single chain antibodies ; scFv ; potato cyst nematodes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests. Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (VL) and heavy chain (VH) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974485
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  • 10
    Electronic Resource
    Electronic Resource
    Identification and management of virulence genes in potato cyst nematodes (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 157-163 
    Details
    ISSN: 1573-8469
    Keywords: Globodera rostochiensis ; G. pallida ; hybridoma ; PCR ; pathotypes ; RAPD ; 2D-gel electrophoresis ; virulence genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Efficient and accurate diagnostic assays are essential for the design and evaluation of control measures of the potato cyst nematodesGlobodera rostochiensis andG. pallida by means of resistance. The hybridoma technology and the polymerase chain reaction (PCR) offer in potential various possibilities to design such diagnostic tests for routine purposes. We set out to devise a refined advisory system based on biochemical assays by using the following stepwise approach. In the early 80's a research program was started to develop an immunoassay to differentiate the two sibling species of potato cyst nematodes. Species specific monoclonal antibodies were raised against nematode proteins which are thermostable, abundant and homologous, and which enable reliable species identification using single eggs. The second step to improve the management of virulence genes is aimed at discriminating groups of populations within a species (‘virulence groups’ or ‘pathotypes’). The concept is that the number of initial populations introduced from South America is limited and that numerous Dutch populations (‘secondary founders’) are closely related by descent. Biochemical characters revealed by two-dimensional gel electrophoresis (2-DGE) of polypeptides, PCR in combination with restriction enzyme digests and RAPD (Random amplified polymorphic DNA) will be used to delineate groups of populations. The final diagnostic assay will be based on PCR. One of the challenges will be to devise a manageable number of primers to recognize all distinct groups. The third research line is aimed at developing a PCR assay based on the virulence genes themselves. Genetic studies showed that virulence inG. rostochiensis towards the H1 resistance gene is inherited at a single locus and is recessive to avirulence. To identify molecular markers linked to the virulence gene, 300 virulent lines were selected via backcrossing the F1 (Aa) with the virulent (aa) parent line. Molecular differences between the parent lines were obtained by 2-DGE, RFLP's (restriction fragment length polymorphisms) and RAPD. Especially RAPD proved to be a valuable technique to construct a linkage map. Screening 80 primers (10-mer) resolved more than 120 markers. RAPD will eventually lead to flanking DNA sequences, which will be used to isolate and characterize the virulence gene. Sequence information of the virulence gene inG. rostochiensis for the H1 resistance gene can be used to devise primers for a PCR assay and may also provide a starting point to isolate other virulence genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974482
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