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efficient agrobacterium tumefaciens-mediated transformation and regeneration of garlic (allium sativum) immature leaf tissue.transgenic garlic (allium sativum) plants have been recovered directly from immature leaf material by selective culture following agrobacterium-mediated transformation. this method involved the use of a binary vector containing the mgfp-er reporter gene and hpt selectable marker, and followed a similar protocol developed previously for the transformation of immature onion embryos. the choice of tissue and post-transformation selection procedure resulted in a large increase in recovery of transge ...201020099065
genetic transformation of carnation (dianthus caryophylus l.).this chapter describes a rapid and efficient protocol for explant preparation and genetic transformation of carnation. node explants from greenhouse-grown plants and leaf explants from in vitro plants are infected with agrobacterium tumefaciens agl0 harboring pkt3 plasmid, consisting of gus and nptii genes. explant preparation is an important factor to obtain the transformed plants. the gus-staining area was located only on the cut end of explants and only explants with a cut end close to the co ...201020099093
validation of a newly developed hexaplex real-time pcr assay for screening for presence of gmos in food, feed and seed.for years, an increasing number and diversity of genetically modified plants has been grown on a commercial scale. the need for detection and identification of these genetically modified organisms (gmos) calls for broad and at the same time flexible high throughput testing methods. here we describe the development and validation of a hexaplex real-time polymerase chain reaction (pcr) screening assay covering more than 100 approved gmos containing at least one of the gmo targets of the assay. the ...201020101506
effect of potato virus y on the nadp-malic enzyme from nicotiana tabacum l.: mrna, expressed protein and activity.the effect of biotic stress induced by viral infection (potato virus y, strain ntn and o) on nadp-malic enzyme (ec 1.1.1.40) in tobacco plants (nicotiana tabacum l., cv. petit havana, sr1) was tested at the transcriptional, translational and activity level. the increase of enzyme activity in infected leaves was correlated with the increased amount of expressed protein and with mrna of cytosolic nadp-me isoform. transcription of the chloroplastic enzyme was not influenced by viral infection. the ...200920111689
[effects of pa-pex 11 gene on penicillin production in penicillium aurantiogriseum].to study the effect of pa-pex 11 gene on penicillin production in pencillium aurantiogriseum.200920112676
s-glycoprotein-like protein regulates defense responses in nicotiana plants against ralstonia solanacearum.rsrga4 (for ralstonia solanacearum-responsive gene a4) encodes a polypeptide similar to s-locus glycoprotein (sgp) from brassica rapa and sgp-like proteins from ipomoea trifida and medicago truncatula. therefore, we designated rsrga4 as ntsglp (for nicotiana tabacum sgp-like protein) and nbsglp (its nicotiana benthamiana ortholog). nbsglp is expressed in root, leaf, petal, gynoecium, and stamen. expression of nbsglp was strongly induced by inoculation with an avirulent strain of r. solanacearum ...201020118275
plasmid pami2 of paracoccus aminophilus jcm 7686 carries n,n-dimethylformamide degradation-related genes whose expression is activated by a luxr family regulator.n,n-dimethylformamide (dmf), a toxic solvent used in the chemical industry, is frequently present in industrial wastes. plasmid pami2 (18.6 kb) of paracoccus aminophilus jcm 7686 carries genetic information which is crucial for methylotrophic growth of this bacterium, using dmf as the sole source of carbon and energy. besides a conserved backbone related to pagk84 of agrobacterium radiobacter k84, pami2 carries a three-gene cluster coding for the protein dmfr, which has sequence similarities to ...201020118371
agroinfiltration reduces aba levels and suppresses pseudomonas syringae-elicited salicylic acid production in nicotiana tabacum.agrobacterium tumefaciens strain gv3101 (pmp90) is widely used in transient gene expression assays, including assays to study pathogen effectors and plant disease resistance mechanisms. however, inoculation of a. tumefaciens gv3101 into nicotiana tabacum (tobacco) leaves prior to infiltration with pathogenic and non-host strains of pseudomonas syringae results in suppression of macroscopic symptoms when compared with leaves pre-treated with a buffer control.201020126459
agrobacterium radiobacter bacteremia in a child with acute lymphoblastic leukemia.agrobacteria are gram-negative tumorigenic plant pathogens that rarely cause infections in humans.201020127221
agrobacterium type iv secretion system and its substrates form helical arrays around the circumference of virulence-induced cells.the genetic transformation of plant cells by agrobacterium tumefaciens results from the transfer of dna and proteins via a specific virulence (vir) -induced type iv secretion system (t4ss). to better understand t4ss function, we analyzed the localization of its structural components and substrates by deconvolution fluorescence microscopy. gfp fusions to t4ss proteins with cytoplasmic tails, virb8 and vird4, or cytoplasmic t4ss substrate proteins, vird2, vire2, and virf, localize in a helical pat ...201020133577
over-expression of superoxide dismutase exhibits lignification of vascular structures in arabidopsis thaliana.the present study demonstrated that over-expression of copper-zinc superoxide dismutase (cu/zn-sod), an important enzyme scavenging reactive oxygen species, improved vascular structures through lignification and imparted tolerance to salt stress (nacl) in arabidopsis thaliana (arabidopsis; accession col-0). transgenic plants of arabidopsis were developed by over-expressing cytosolic cu/zn-sod from potentilla atrosanguinea under camv35s promoter via agrobacterium tumefaciens mediated transformati ...201020138686
transgenic leucaena leucocephala expressing the rhizobium gene pyda encoding a meta-cleavage dioxygenase shows reduced mimosine content.the use of the tree-legume leucaena leucocephala (leucaena), which contains high levels of proteins in its foliage, is limited due to the presence of the toxic free amino acid mimosine. the goal of this research was to develop transgenic leucaena with reduced mimosine content. two genes, pyda and pydb, encoding a meta-cleavage dioxygenase (ec 1.13.11.2) and a pyruvate hydrolase (ec 3.7.1.6), respectively, from the mimosine-degrading leucaena symbiont rhizobium sp. strain tal1145, were used to tr ...201020138776
biochemical characterization of three putative atpases from a new type iv secretion system of aeromonas veronii plasmid pac3249a.type four secretion systems (tfss) are bacterial macromolecular transport systems responsible for transfer of various substrates such as proteins, dna or protein-dna complexes. tfsss encode two or three atpases generating energy for the secretion process. these enzymes exhibit highest sequence conservation among type four secretion components.201020144229
transformation of the plant kalanchoë daigremontiana using agrobacterium tumefaciens.kalanchoë daigremontiana can be stably transformed using the agrobacterium tumefaciens-mediated t-dna transfer method, as described here. sterilized plant tissue is cocultivated with an a. tumefaciens suspension, transformants are selected and the shoots are grown in rooting medium and then in soil. plant phenotypes can be examined approximately 3 mo after transfer of plants to soil.200920147048
agrobacterium tumefaciens-mediated transformation of cleome gynandra l., a c(4) dicotyledon that is closely related to arabidopsis thaliana.in leaves of most c(4) plants, the biochemistry of photosynthesis is partitioned between mesophyll and bundle sheath cells. in addition, their cell biology and development also differs from that in c(3) plants. we have a poor understanding of the mechanisms that generate the cell-specific accumulation of proteins used in the c(4) pathway, and there are few genes that have been shown to be important for the cell biology and development of c(4) leaves. to facilitate functional analysis of c(4) pho ...201020150516
new insights into an old story: agrobacterium-induced tumour formation in plants by plant transformation.agrobacterium tumefaciens causes tumour formation in plants. plant signals induce in the bacteria the expression of a range of virulence (vir) proteins and the formation of a type iv secretion system (t4ss). on attachment to plant cells, a transfer dna (t-dna) and vir proteins are imported into the host cells through the bacterial t4ss. through interaction with a number of host proteins, the vir proteins suppress the host innate immune system and support the transfer, nuclear targeting, and inte ...201020150897
molecular genetic analysis of the orsellinic acid/f9775 gene cluster of aspergillus nidulans.f-9775a and f-9775b are cathepsin k inhibitors that arise from a chromatin remodelling deletant strain of aspergillus nidulans. a polyketide synthase gene has been determined to be responsible for their formation and for the simpler, archetypical polyketide orsellinic acid. we have discovered simple culture conditions that result in the production of the three compounds, and this facilitates analysis of the genes responsible for their synthesis. we have now analysed the f9775/orsellinic acid gen ...201020174687
phylogenomics reveals a diverse rickettsiales type iv secretion system.with an obligate intracellular lifestyle, alphaproteobacteria of the order rickettsiales have inextricably coevolved with their various eukaryotic hosts, resulting in small, reductive genomes and strict dependency on host resources. unsurprisingly, large portions of rickettsiales genomes encode proteins involved in transport and secretion. one particular transporter that has garnered recent attention from researchers is the type iv secretion system (t4ss). homologous to the well-studied archetyp ...201020176788
protocol: a rapid and economical procedure for purification of plasmid or plant dna with diverse applications in plant biology.abstract: research in plant molecular biology involves dna purification on a daily basis. although different commercial kits enable convenient extraction of high-quality dna from e. coli cells, pcr and agarose gel samples as well as plant tissues, each kit is designed for a particular type of dna extraction work, and the cost of purchasing these kits over a long run can be considerable. furthermore, a simple method for the isolation of binary plasmid from agrobacterium tumefaciens cells with sat ...201020180960
resistance to crown gall disease in transgenic grapevine rootstocks containing truncated vire2 of agrobacterium.a truncated form of the ti-plasmid vire2 gene from agrobacterium tumefaciens strains c58 and a6, and a. vitis strain cg450 was transferred and expressed in somatic embryos of grapevine rootstocks 110 richter (vitis rupestris × v. berlandieri), 3309 couderc (v. rupestris × v. riparia) and teleki 5c (v. berlandieri × v. riparia) via agrobacterium-mediated transformation to confer resistance to crown gall disease. transformation was confirmed in 98% of the 322 lines by enzyme-linked immunosorbent a ...201020182792
identification of a dehydrogenase required for lactose metabolism in caulobacter crescentus.caulobacter crescentus, which thrives in freshwater environments with low nutrient levels, serves as a model system for studying bacterial cell cycle regulation and organelle development. we examined its ability to utilize lactose (i) to gain insight into the metabolic capacities of oligotrophic bacteria and (ii) to obtain an additional genetic tool for studying this model organism, aiming to eliminate the basal enzymatic activity that hydrolyzes the chromogenic substrate 5-bromo-4-chloro-3-indo ...201020190087
[generation of sugar beet transgenic plants expressing bar gene].the parameters of transformation using agrobacterium tumefaciens eha 105 for 5 domestic sorts and lines of sugar beet (beta vulgaris l. var. saccharifera (alef) krass) were optimized. the system of transgenic tissue selection based on resistance to phosphinothricin, allowing to avoid the appearing of chimeric shoots among initial transformants was developed. the transgenic plants of sugar beet sorts ramonskaya single seed 47, l'govskaya single seed 52 and rms 73, and lbo 17 and lbo 19 lines expr ...201020198924
selectable marker elimination in the t0 generation by agrobacterium-mediated co-transformation involving mungbean yellow mosaic virus trap as a non-conditional negative selectable marker and bar for transient positive selection.transient selection involving the bar gene and non-conditional negative selection against stable t-dna integration through the use of the mungbean yellow mosaic virus (mymv) transcriptional activator protein gene (trap) were used in a novel co-transformation strategy to generate selectable marker gene (smg)-eliminated transgenic tobacco plants in the t(0) generation itself. two compatible binary plasmids, pcam-bar-trap-gus harbouring bar as an smg and the mymv trap gene as a non-conditional nega ...201020204372
effect of limited oxygen supply on coenzyme q(10) production and its relation to limited electron transfer and oxidative stress in rhizobium radiobacter t6102.coenzyme q(10) (coq(10)) productions from rhizobium radiobacter t6102 were monitored under various oxygen supply conditions by controlling the agitation speeds, aeration rates, and dissolved oxygen levels. as the results, the coq(10) production was enhanced by limited oxygen supply. to investigate whether the coq(10) production is associated with its physiological functions of electron carrier and antioxidant, the effects of sodium azide and hydrogen peroxide on the coq(10) production were studi ...201020208439
cloning of a nacl-induced fructose-1, 6-diphosphate aldolase cdna from dunaliella salina and its expression in tobacco.using rapid amplification of cdna ends (race) technique, the full-length cdna encoding a nacl-induced fructose-1, 6-diphosphate aldolase (dsaldp) was obtained. it was shown that the dsaldp had a relatively high homology (66%-73%) to chloroplast fructose-1, 6-diphosphate aldolase (aldp) in many plants according to their amino acid sequences. the phylogenetic analysis further confirmed that aldp in alga is the nearest to dsaldp. as to its expression pattern, dsaldp was de novo synthesized by nacl ...200320213361
an efficient method for the production of marker-free transgenic plants of peanut (arachis hypogaea l.).recombinant genes conferring resistance to antibiotics or herbicides are widely used as selectable markers in plant transformation for selecting the primary transgenic events. however, these become redundant once the transgenic plants have been developed and identified. although, there is no evidence that the selectable marker genes are unsafe for consumers and the environment, it would be desirable if the marker genes can be eliminated from the final transgenic events. the availability of effic ...201020217416
a method to confer salinity stress tolerance to plants by helicase overexpression.high salinity stress adversely affects plant growth and limits agricultural production worldwide. to minimize these losses it is essential to develop stress-tolerant plants. several genes, including the genes encoding for helicases, are induced in response to salinity stress. helicases are ubiquitous motor enzymes that catalyze the unwinding of energetically stable duplex dna (dna helicases) or duplex rna secondary structures (rna helicases) in an atp-dependent manner. helicase members of dead-b ...201020225163
functional characterization and manipulation of the apicidin biosynthetic pathway in fusarium semitectum.apicidin is a cyclic tetrapeptide produced by certain isolates of fusarium semitectum and has been shown to inhibit apicomplexan histone deacetylase. an apicidin-producing strain (kctc16676) of the filamentous fungus was mutated using an agrobacterium tumefaciens-mediated transformation, resulting in 24 apicidin-deficient mutants. three of the mutants had a t-dna insertion in a gene that encodes a non-ribosomal peptide synthetase (nrps). results of sequence, expression, and gene deletion analyse ...201020233305
in situ raman and x-ray spectroscopies to monitor microbial activities under high hydrostatic pressure.until recently, monitoring of cells and cellular activities at high hydrostatic pressure (hhp) was mainly limited to ex situ observations. samples were analyzed prior to and following the depressurization step to evaluate the effect of the pressure treatment. such ex situ measurements have several drawbacks: (i) it does not allow for kinetic measurements and (ii) the depressurization step often leads to artifactual measurements. here, we describe recent advances in diamond anvil cell (dac) techn ...201020233376
phosphomannose isomerase gene for selection in lettuce (lactuca sativa l.) transformation.a positive selection system using phosphomannose isomerase was employed for agrobacterium tumefaciens mediated transformation of lettuce (lactuca sativa l. var. 'achát'). it was shown that the mannose-based selection system works very well with the lettuce genotype used, reaching up to 25% transformation efficiency on the medium with 20 g/l mannose and 20 g/l sucrose. the best transformation efficacy with the commonly-used kanamycin at 100 mg/l as a selection agent was 21%. southern blot analyse ...201020234883
electroporation and agrobacterium-mediated spore transformation.genetic transformation is a key technology in modern fungal research. most commonly, protoplasts are transformed using the polyethylene glycol-mediated transformation protocols. because protoplasts are generated by treatment of mycelia with a crude enzyme preparation, the results tend to be inconsistent. furthermore, some species cannot be transformed by this method. electroporation (ep) and agrobacterium tumefaciens-mediated transformation (amt) are two alternative methods. these methods allow ...201020238258
[not available]. 194720268193
the green fluorescent protein as an efficient selection marker for agrobacterium tumefaciens-mediated transformation in hevea brasiliensis (müll. arg).an efficient genetic transformation procedure using a recombinant green fluorescent protein (gfp) has been developed in hevea brasiliensis clone pb260. transformation experiments have been performed using an agrobacterium tumefaciens binary vector harbouring both uida and s65t-gfp reporter genes in order to compare selection methods using glucuronidase assay (gus activity) and paromomycin resistance, gfp activity and paromomycin resistance, or gfp activity only. at transient level, the number of ...201020306052
using high competent shoot apical meristems of cockscomb as explants for studying function of asymmetric leaves2-like11 (asl11) gene of arabidopsis.though shoot apical meristems (sams) commonly exhibit low or no competence for transformation, the potent regeneration of this tissue merits further research. especially, when shoot regeneration is recalcitrant using other tissues as explants, sam probably is an excellent selection. in cockscomb plants, using sams from seedlings obtained from ms medium with 0.5 mg l(-1) 6-ba as explants, high frequency of transformation (approximate 20%) is obtained; whereas control sams performed poorly for tra ...201020306306
light-induced activation of bacterial phytochrome agp1 monitored by static and time-resolved ftir spectroscopy.phytochromes, which regulate many biological processes in plants, bacteria, and fungi, can exist in two stable states, pr and pfr, that can be interconverted by light, via a number of intermediates such as meta-rc. herein we employ ftir spectroscopy to study the pr-to-pfr conversion of the bacteriophytochrome agp1 from agrobacterium tumefaciens. static ftir pfr/pr and meta-rc/pr difference spectra are disentangled in terms of cofactor and protein structural changes. guided by dft calculations on ...201020333618
induction of chromosomal inversion by integration of t-dna in the rice genome.transfer dna (t-dna) of agrobacterium tumefaciens integration in the plant genome may lead to rearrangements of host plant chromosomal fragments, including inversions. however, there is very little information concerning the inversion. the present study reports a transgenic rice line selected from a t-dna tagged population, which displays a semi-dwarf phenotype. molecular analysis of this mutant indicated an insertion of two tandem copies of t-dna into a locus on the rice genome in a head to tai ...201020347828
agrobacterium tumefaciens type iv secretion protein virb3 is an inner membrane protein and requires virb4, virb7, and virb8 for stabilization.agrobacterium tumefaciens virb proteins assemble a type iv secretion apparatus and a t-pilus for secretion of dna and proteins into plant cells. the pilin-like protein virb3, a membrane protein of unknown topology, is required for the assembly of the t-pilus and for t-dna secretion. using phoa and green fluorescent protein (gfp) as periplasmic and cytoplasmic reporters, respectively, we demonstrate that virb3 contains two membrane-spanning domains and that both the n and c termini of the protein ...201020348257
increasing anthraquinone production by overexpression of 1-deoxy-d: -xylulose-5-phosphate synthase in transgenic cell suspension cultures of morinda citrifolia.a morinda citrifolia cell line was obtained by overexpresion of 1-deoxy-d: -xylulose 5-phosphate synthase (dxs) from catharanthus roseus, a key enzyme of the metabolic pathway of anthraquinones (aqs). this cell line increased aqs production by about 24% compared to the control cell line. this transgenic cell line which carries dxs cdna isolated from catharanthus roseus, was achieved by direct transformation of cell suspension cultures of m. citrifolia using a hypervirulent agrobacterium tumefaci ...201020349334
agrobacterium tumefaciens-mediated transformation of maize endosperm as a tool to study endosperm cell biology.developing maize (zea mays) endosperms can be excised from the maternal tissues and undergo tissue/cell-type differentiation under in vitro conditions. we have developed a method to transform in vitro-grown endosperms using agrobacterium tumefaciens and standard binary vectors. we show that both aleurone and starchy endosperm cells can be successfully transformed using a short cocultivation with a. tumefaciens cells. the highest transformation rates were obtained with the a. tumefaciens eha101 s ...201020357137
[clinical and microbiological characteristics of rhizobium radiobacter infectious: six cases report and literature review].to explore the clinical and microbiological characteristics of rhizobium radiobacter infection, and therefore to provide information for the prevention and treatment of the disease.201020367947
sreb, a gata transcription factor that directs disparate fates in blastomyces dermatitidis including morphogenesis and siderophore biosynthesis.blastomyces dermatitidis belongs to a group of human pathogenic fungi that exhibit thermal dimorphism. at 22 degrees c, these fungi grow as mold that produce conidia or infectious particles, whereas at 37 degrees c they convert to budding yeast. the ability to switch between these forms is essential for virulence in mammals and may enable these organisms to survive in the soil. to identify genes that regulate this phase transition, we used agrobacterium tumefaciens to mutagenize b. dermatitidis ...201020368971
light-induced conformational changes of the chromophore and the protein in phytochromes: bacterial phytochromes as model systems.recombinant phytochromes agp1 and agp2 from agrobacterium tumefaciens are used as model phytochromes for biochemical and biophysical studies. in biliverdin binding phytochromes the site for covalent attachment of the chromophore lies in the n-terminal region of the protein, different from plant phytochromes. the issue which stereochemistry the chromophore adopts in the so-called pr and pfr forms is addressed by using a series of locked chromophores which form spectrally characteristic adducts wi ...201020373318
the grape microvine - a model system for rapid forward and reverse genetics of grapevines.a grapevine model system is described that is suitable for rapid forward and reverse genetic studies in small controlled environments. it is based on the vvgai1 mutant allele that confers a dwarf stature, short generation cycles and continuous flowering ('microvine'). black and white berry microvine genotypes were developed that can be transformed by agrobacterium tumefaciens. near-homozygous lines were created for efficient bi-allelic single nucleotide polymorphism (snp) marker mapping and muta ...201020374531
improvement of coenzyme q(10) production by increasing the nadh/nad(+) ratio in agrobacterium tumefaciens.previously screened coq(10)-overproducing agrobacterium tumefaciens a603-35 showed a relatively high nadh/nad(+) ratio (1.1), as compared to parental strain c58 (0.2) when we increased the expression levels of nadh-generating enzymes. also, the intracellular nadh/nad(+) ratio showed a positive correlation with the coq(10) content in a603-35. overexpression of glyceraldehyde 3-phosphate dehydrogenase in a603-35 shifted the nadh/nad(+) ratio at 48 h from 0.8 to 1.2, and thus the coq(10) content in ...201020378960
[construction of t-dna insertion mutants of botrytis cinerea via agrobacterium tumefaciens mediated transformation and sequence analysis of insertion site].construction of botrytis cinerea t-dna insertion mutant library through agrobacterium-mediated transformation and for further understanding of the molecular mechanisms underlying the pathogenicity of botrytis cinerea.201020387458
novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of xanthomonas fuscans subsp. aurantifolii.citrus canker is a disease that has severe economic impact on the citrus industry worldwide. there are three types of canker, called a, b, and c. the three types have different phenotypes and affect different citrus species. the causative agent for type a is xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. xanthomonas fuscans subsp. aurantifolii strain b causes canker b and xanthomonas fuscans subsp. aurantifolii strain c causes canker c.201020388224
monomethylamine as a nitrogen source for a nonmethylotrophic bacterium, agrobacterium tumefaciens.monomethylamine can be used by nonmethylotrophs as a sole nitrogen source but not as a carbon source; however, little is known about the genes and enzymes involved. the gamma-glutamylmethylamide/n-methylglutamate pathway for monomethylamine utilization by methylotrophs has recently been resolved. we have identified genes encoding key enzymes of this pathway in nonmethylotrophs (e.g., agrobacterium tumefaciens) and demonstrated that this pathway is also involved in the utilization of monomethylam ...201020400554
a role for psk signaling in wounding and microbial interactions in arabidopsis.psk-alpha is a disulfated peptide that acts as a growth factor in plants. psk-alpha is derived from preproproteins which are encoded by five psk precursor genes in arabidopsis thaliana (l.) heynh and is perceived by leucine-rich repeat receptor kinases. arabidopsis has two psk receptor genes, pskr1 and pskr2. although ligand and receptor are well characterized, the biological functions of psk signaling are not well understood. using reporter lines and receptor knockout mutants of arabidopsis, a ...201020403122
the batr/bats two-component regulatory system controls the adaptive response of bartonella henselae during human endothelial cell infection.here, we report the first comprehensive study of bartonella henselae gene expression during infection of human endothelial cells. expression of the main cluster of upregulated genes, comprising the virb type iv secretion system and its secreted protein substrates, is shown to be under the positive control of the transcriptional regulator batr. we demonstrate binding of batr to the promoters of the virb operon and a substrate-encoding gene and provide biochemical evidence that batr and bats const ...201020418395
2-de proteome maps for the leaf apoplast of nicotiana benthamiana.we provide 2-d gel reference maps for the apoplastic proteome of nicotiana benthamiana leaves infiltrated or not with the bacterial gene vector agrobacterium tumefaciens. about 90 proteins were analyzed by lc-ms/ms for identification and function assignment. we show, overall, an effective response of the plant to agroinfiltration involving a specific, cell wall maintenance-independent up-regulation of defense protein secretion. the proteome maps described should be a useful tool for systemic stu ...201020422621
the small heat-shock protein hspl is a virb8 chaperone promoting type iv secretion-mediated dna transfer.agrobacterium tumefaciens is a plant pathogen that utilizes a type iv secretion system (t4ss) to transfer dna and effector proteins into host cells. in this study we discovered that an alpha-crystallin type small heat-shock protein (alpha-hsp), hspl, is a molecular chaperone for virb8, a t4ss assembly factor. hspl is a typical alpha-hsp capable of protecting the heat-labile model substrate citrate synthase from thermal aggregation. it forms oligomers in a concentration-dependent manner in vitro. ...201020427270
an efficient in vitro-inoculation method for tomato yellow leaf curl virus.tomato yellow leaf curl virus (tylcv) is a member of the family geminiviridae, genus begomovirus. to test the infectivity of tylcv in tomato plants, an improved protocol for inoculation of in vitro-cultured tomato plants was developed.201020429892
the application of the yeast n-acetyltransferase mpr1 gene and the proline analogue l-azetidine-2-carboxylic acid as a selectable marker system for plant transformation.the yeast n-acetyltransferase mpr1 gene has previously been shown to confer resistance to the toxic proline analogue azetidine-2-carboxylic acid (a2c) in yeast and transgenic tobacco. here experiments were carried out to determine if mpr1 and a2c can work as a selectable marker system for plant transformation. the mpr1 gene was inserted into a binary vector under the control of the cauliflower mosaic virus 35s promoter and nopaline synthase terminator, and transformed into tobacco via the agroba ...201020430752
proteomic and transcriptomic characterization of a virulence-deficient phosphatidylcholine-negative agrobacterium tumefaciens mutant.phosphatidylcholine (pc) is the most abundant phospholipid in eukaryotic membranes, whereas only a limited number of bacteria are able to synthesize pc. intriguingly, many of the bacteria with pc-containing membranes interact with eukaryotic hosts. pc is one of the major membrane lipids in the phytopathogenic bacterium agrobacterium tumefaciens. the presence of pc is critical for diverse cellular processes like motility, biofilm formation, stress resistance, and virulence. the exact role of pc i ...201020437057
rhizobium borbori sp. nov., aniline-degrading bacteria isolated from activated sludge.three aniline-degrading bacteria, strains dn316(t), dn316-1 and dn365, were isolated from activated sludge. according to 16s rrna gene sequence-based phylogenetic analysis, the isolates belonged to the genus rhizobium, with rhizobium ( = agrobacterium) radiobacter lmg 140(t) as the closest relative, with 96.5 % sequence similarity. phylogenetic analysis of the representative strain dn316(t) using sequences of the glna, thrc and reca genes and the 16s-23s intergenic spacer region confirmed the ph ...201120453105
mutation of outer-shell residues modulates metal ion co-ordination strength in a metalloenzyme.the metal ion co-ordination sites of many metalloproteins have been characterized by a variety of spectroscopic techniques and small-molecule model systems, revealing many important insights into the structural determinants of metal ion co-ordination. however, our understanding of this fundamentally and practically important phenomenon remains frustratingly simplistic; in many proteins it is essentially impossible to predict metal ion specificity and the effects of remote 'outer-shell' residues ...201020459397
expression of the fluorescent proteins dsred and egfp to visualize early events of colonization of the chickpea blight fungus ascochyta rabiei.ascochyta blight caused by the ascomycete fungus ascochyta rabiei, is a major biotic constraint of chickpea (cicer arietinum l.), resulting in disastrous crop losses worldwide. to study early stages of development and pathogenic mechanisms of the fungus, two binary vectors for the constitutive expression of the red fluorescent protein (dsred-express) and the green fluorescent protein (egfp1) were constructed. furthermore, we have developed an improved and highly reproducible agrobacterium tumefa ...201020461519
an improved agrobacterium-mediated transformation system for the functional genetic analysis of penicillium marneffei.we have developed an improved agrobacterium-mediated transformation (amt) system for the functional genetic analysis of penicillium marneffei, a thermally dimorphic, human pathogenic fungus. our amt protocol included the use of conidia or pre-germinated conidia of p. marneffei as the host recipient for t-dna from agrobacterium tumefaciens and co-cultivation at 28°c for 36 hours. bleomycin-resistant transformants were selected as yeast-like colonies following incubation at 37°c. the efficiency of ...201020465521
novel functions of stomatal cytokinesis-defective 1 (scd1) in innate immune responses against bacteria.eukaryotes employ complex immune mechanisms for protection against microbial pathogens. here, we identified scd1 (stomatal cytokinesis-defective 1), previously implicated in growth and development through its role in cytokinesis and polarized cell expansion (falbel, t. g., koch, l. m., nadeau, j. a., segui-simarro, j. m., sack, f. d., and bednarek, s. y. (2003) development 130, 4011-4024) as a novel component of innate immunity. in arabidopsis, scd1 is a unique gene encoding for the only protein ...201020472560
vip1: linking agrobacterium-mediated transformation to plant immunity?agrobacterium tumefaciens is the most efficient vehicle used today for the production of transgenic plants and plays an essential role in basic scientific research and in agricultural biotechnology. previously, plant vire2-interacting protein 1 (vip1) was shown to play a role in agrobacterium-mediated transformation. recent reports demonstrate that vip1, as one of the bzip transcription factors, is also involved in plant immunity responses. agrobacterium is able to activate and abuse vip1 for tr ...201020473505
enhancing the virulence of paecilomyces lilacinus against meloidogyne incognita eggs by overexpression of a serine protease.to enhance the virulence of paecilomyces lilacinus against meloidogyne incognita eggs, a serine protease was overexpressed in p. lilacinus 9410 (pl9410). the cdna of a mature serine protease gene was cloned from pl9410 and integrated into the genomes of pl9410 transformants through agrobacterium tumefaciens-mediated transformation. our results confirmed that the serine protease gene was overexpressed at the transcriptional level, and that the serine protease activities were enhanced in the trans ...201020473777
a novel cold-active and alkali-stable β-glucosidase gene isolated from the marine bacterium martelella mediterranea.a β-glucosidase gene designated gluc3m was cloned through construction of a genomic library of martelella mediterranea 2928. the gluc3m consisted of 2,496 bp and encoded a peptide of 832 amino acids that shared the greatest amino acid similarity (59%) with a β-glucosidase of family 3 glycoside hydrolase from agrobacterium radiobacter k84. the optimum reaction temperature and ph of gluc3m were 45 °c and 8.0, respectively. the k (m) and v (max) for p-nitrophenyl-β-d: -glucopyranoside were 0.18 mg/ ...201020503105
fusarium wilt (fusarium oxysporum f. sp. vasinfectum) genes expressed during infection of cotton (gossypium hirsutum)dagger.summary we sought to identify fusarium oxysporum f. sp. vasinfectum (fov) genes that may be associated with pathogenicity. initially we utilized microarray and q-pcr technology to identify fov genes expressed in root and hypocotyl tissues during a compatible infection of cotton. we identified 218 fungal clones representing 174 fov non-redundant genes as expressed in planta. the majority of the expressed sequences were expressed in infected roots, with only six genes detected in hypocotyl tissue. ...200620507430
transgenic arabidopsis plants expressing agrobacterium tumefaciens vird2 protein are less susceptible to agrobacterium transformation.summary agrobacterium tumefaciens causes crown gall disease on many plant species and can result in considerable economic losses. here we report a new strategy to control crown gall disease by over-expressing agrobacterium tumefaciens vird2 protein in plants. transgenic arabidopsis plants over-expressing vird2 from constitutive or wound-inducible promoters are less susceptible to agrobacterium-mediated transformation. additionally, the transient introduction of an a. tumefaciens vird2 gene in to ...200620507462
agroinfection-based high-throughput screening reveals specific recognition of inf elicitins in solanum.summary we adapted and optimized the use of the agrobacterium tumefaciens binary pvx expression system (pvx agroinfection) to screen solanum plants for response to pathogen elicitors and applied the assay to identify a total of 11 clones of solanum huancabambense and solanum microdontum, out of 31 species tested, that respond to the elicitins inf1, inf2a and inf2b of phytophthora infestans. prior to this study, response to inf elicitins was only known in nicotiana spp. within the solanaceae. the ...200620507464
the tobacco ntann12 gene, encoding an annexin, is induced upon rhodoccocus fascians infection and during leafy gall development.summary annexins are calcium-binding proteins that have been associated in plants with different biological processes such as responses to abiotic stress and early nodulation stages. until now, the implication of annexins during plant-pathogen interactions has not been reported. here, a novel plant annexin gene induced in tobacco by-2 cell suspension cultures infected with the phytopathogenic bacterium rhodococcus fascians (strain d188) has been identified. expression of this gene, called ntann1 ...200720507490
crystal structure of exotype alginate lyase atu3025 from agrobacterium tumefaciens.alginate, a major component of the cell wall matrix in brown seaweeds, is degraded by alginate lyases through a beta-elimination reaction. almost all alginate lyases act endolytically on substrate, thereby yielding unsaturated oligouronic acids having 4-deoxy-l-erythro-hex-4-enepyranosyluronic acid at the nonreducing end. in contrast, agrobacterium tumefaciens alginate lyase atu3025, a member of polysaccharide lyase family 15, acts on alginate polysaccharides and oligosaccharides exolytically an ...201020507980
rnai-mediated suppression of the phenylalanine ammonia-lyase gene in salvia miltiorrhiza causes abnormal phenotypes and a reduction in rosmarinic acid biosynthesis.medicinal salvia miltiorrhiza contains two main groups of active pharmaceutical ingredients: lipid-soluble tanshinones and water-soluble phenolic acids, including rosmarinic acid and salvianolic acid b. phenylalanine ammonia-lyase (pal) catalyzes the first step in the phenylpropanoid pathway and is assumed to be closely related to the accumulation of rosmarinic acid and its derivatives. we selected a 217-bp fragment, located at the 3' end of the coding region of pal1, to establish an rna interfe ...201120509039
rapid and efficient identification of agrobacterium species by reca allele analysis: agrobacterium reca diversity.the analysis of housekeeping reca gene sequences from 138 strains from 13 species or genomic species of agrobacterium, nine being biovar 1 genomospecies, and the others agrobacterium larrymoorei, agrobacterium rubi, agrobacterium sp. ncppb 1650, and agrobacterium vitis and one "former" agrobacterium species, rhizobium rhizogenes, led to the identification of 50 different reca alleles and to a clear delineation of the 14 species or genomospecies entirely consistent with that obtained by amplified ...201020521039
beauvericin from the endophytic fungus, fusarium redolens, isolated from dioscorea zingiberensis and its antibacterial activity.from the fungal endophyte, fusarium redolens dzf2, isolated from rhizomes of the chinese medicinal plant dioscorea zingiberensis, beauvericin was obtained by tlc in combination with bioautographic antibacterial assay. the compound was identified by spectroscopic and physicochemical means. the median effective inhibitory concentration (ic50) values of beauvericin against six test bacteria (bacillus subtilis, staphylococcus haemolyticus, pseudomonas lachrymans, agrobacterium tumefaciens, escherich ...201020521553
expression of mycobacterium leprae hsp65 in tobacco and its effectiveness as an oral treatment in adjuvant-induced arthritis.transgenic plants are able to express molecules with antigenic properties. in recent years, this has led the pharmaceutical industry to use plants as alternative systems for the production of recombinant proteins. plant-produced recombinant proteins can have important applications in therapeutics, such as in the treatment of rheumatoid arthritis (ra). in this study, the mycobacterial hsp65 protein expressed in tobacco plants was found to be effective as a treatment for adjuvant-induced arthritis ...201120526808
isolation and antisense suppression of flavonoid 3', 5'-hydroxylase modifies flower pigments and colour in cyclamen.cyclamen is a popular and economically significant pot plant crop in several countries. molecular breeding technologies provide opportunities to metabolically engineer the well-characterized flavonoid biosynthetic pathway for altered anthocyanin profile and hence the colour of the flower. previously we reported on a genetic transformation system for cyclamen. our aim in this study was to change pigment profiles and flower colours in cyclamen through the suppression of flavonoid 3', 5'-hydroxylas ...201020540805
floral-dip transformation of arabidopsis thaliana to examine ptso2::beta-glucuronidase reporter gene expression.the ability to introduce foreign genes into an organism is the foundation for modern biology and biotechnology. in the model flowering plant arabidopsis thaliana, the floral-dip transformation method has replaced all previous methods because of its simplicity, efficiency, and low cost. specifically, shoots of young flowering arabidopsis plants are dipped in a solution of agrobacterium tumefaciens carrying specific plasmid constructs. after dipping, the plants are returned to normal growth and yi ...201020548285
stable integration and expression of wasabi defensin gene in "egusi" melon (colocynthis citrullus l.) confers resistance to fusarium wilt and alternaria leaf spot.production of "egusi" melon (colocynthis citrullus l.) in west africa is limited by fungal diseases, such as alternaria leaf spot and fusarium wilt. in order to engineer "egusi" resistant to these diseases, cotyledonary explants of two "egusi" genotypes, 'ejagham' and nhc1-130, were transformed with agrobacterium tumefaciens strain eha101 harbouring wasabi defensin gene (isolated from wasabia japonica l.) in a binary vector pekh1. after co-cultivation for 3 days, infected explants were transferr ...201020552202
plant-expressed recombinant mountain cedar allergen jun a 1 is allergenic and has limited pectate lyase activity.mountain cedar (juniperus ashei) pollen commonly causes a winter time allergic rhinitis in the central usa. jun a 1 is the dominant allergenic protein, but biologically active recombinant jun a 1 has not been successfully expressed, despite numerous attempts with several expression systems.201020559000
agrobacterium-mediated t-dna transfer and integration into the chromosome of streptomyces lividans.summary agrobacterium tumefaciens is the prototype of a prokaryotic organism transmitting dna across natural kingdom barriers into higher cells. in nature, a specific segment (t-dna) of the resident ti plasmid is transferred from this bacterium into plant cells and integrated into the plant cell genome. expression of the integrated oncogenes contained in the t-dna results in the tumour disease known as crown gall. besides plants, the range of transformable recipients is broad and includes fungi ...200220569318
characterization of oncogene-silenced transgenic plants: implications for agrobacterium biology and post-transcriptional gene silencing.summary agrobacterium tumefaciens tumorigenesis is initiated by the horizontal transfer of a suite of oncogenes that alter hormone synthesis and sensitivity in infected plant cells. transgenic plants silenced for the iaam and ipt oncogenes are highly recalcitrant to tumorigenesis, and present a unique resource to elucidate fundamental questions related to agrobacterium biology and post-transcriptional gene silencing (ptgs). the oncogene-silenced transgenic tomato line 01/6 was used to characteri ...200320569363
agrobacterium tumefaciens mediated transformation of the oomycete plant pathogen phytophthora infestans.summary agrobacterium tumefaciens is widely used for plant dna transformation and, more recently, has also been used to transform yeast and filamentous fungi. here we present a protocol for agrobacterium-mediated dna transformation of the oomycete phytophthora infestans, the causal agent of potato late blight. binary t-dna vectors containing neomycin phosphotransferase (npt) and beta-glucuronidase (gus) fused to oomycete transcriptional regulatory sequences were constructed. seven days of co-cul ...200320569405
backbone-free transformation of barrel medic (medicago truncatula) with a medicago-derived transfer dna.in the present work, agrobacterium tumefaciens-mediated genetic transformation of the model legume medicago truncatula gaertn. (barrel medic) was carried out using the psim843 vector that contains a medicago-derived transfer dna, delineated by a 25-bp sequence homologous to bacterial t-dna borders. the transfer dna contains an expression cassette for the nptii (neomycin phosphotransferase) gene and is flanked by an expression cassette for the backbone integration marker gene ipt (isopentenyl tra ...201020571798
from host recognition to t-dna integration: the function of bacterial and plant genes in the agrobacterium-plant cell interaction.abstract agrobacterium tumefaciens and its related species, a. rhizogenes and a. vitis, are the only known bacterial pathogens which 'genetically invade' host plants and stably integrate part of their genetic material into the host cell genome. thus, a. tumefaciens has evolved as a major tool for plant genetic engineering. furthermore, this unique process of interkingdom dna transfer has been utilized as a model system for studies of its underlying biological events, such as intercellular signal ...200020572967
nematodes as vectors to introduce agrobacterium into plant roots.summary a fast plant promoter test was developed by means of a nematode to transfer agrobacterium tumefaciens into plant roots. two-week-old arabidopsis thaliana (l.) heynh. plants were transferred to infection medium. meloidogyne incognita or heterodera schachtii juveniles were mixed with the agrobacterium strain that harboured the binary vector, and this mixture was used for plant inoculation. during migration of the nematode and establishment of the feeding site inside the roots, the t-dna wa ...200020572986
type iii secretion and in planta recognition of the xanthomonas avirulence proteins avrbs1 and avrbst.summary the hrp gene cluster of the plant pathogen xanthomonas campestris pv. vesicatoria (xcv) encodes a type iii secretion system required for the delivery of virulence and avirulence proteins into the plant. some of these effector proteins, e.g. avrbs1 and avrbst, are recognized by pepper plants carrying corresponding resistance genes, triggering the hypersensitive reaction (hr). in this study, epitope tagged avrbs1 and avrbst proteins were detected in culture supernatants only in the presenc ...200120573017
agrobacterium tumefaciens exor represses succinoglycan biosynthesis and is required for biofilm formation and motility.the ubiquitous plant pathogen agrobacterium tumefaciens attaches efficiently to plant tissues and abiotic surfaces and can form complex biofilms. a genetic screen for mutants unable to form biofilms on pvc identified disruptions in a homologue of the exor gene. exor is a predicted periplasmic protein, originally identified in sinorhizobium meliloti, but widely conserved among alphaproteobacteria. disruptions in the a. tumefaciens exor gene result in severely compromised attachment to abiotic sur ...201020576688
gene transfer into solanum tuberosum via rhizobium spp.agrobacterium tumefaciens-mediated transformation (atmt) is the preferred technique for gene transfer into crops. a major disadvantage of the technology remains the complexity of the patent landscape that surrounds atmt which restricts its use for commercial applications. an alternative system has been described (broothaerts et al. in nature 433:629-633, 2005) detailing the propensity of three rhizobia to transform the model crop arabidopsis thaliana, the non-food crop nicotiana tabacum and, at ...201120582626
rhizobium rosettiformans sp. nov., isolated from a hexachlorocyclohexane dump site, and reclassification of blastobacter aggregatus hirsch and muller 1986 as rhizobium aggregatum comb. nov.a gram-negative, rod-shaped, motile, aerobic bacterial strain, w3(t), was isolated from hexachlorocyclohexane (hch)-contaminated groundwater from lucknow, india, and its taxonomic position was determined using a polyphasic approach. strain w3(t) shared highest 16s rrna gene sequence similarity of 97.8 % with rhizobium selenitireducens b1(t), followed by rhizobium daejeonense l61(t) (97.7 %), rhizobium radiobacter atcc 19358(t) (97.5 %) and blastobacter aggregatus ifam 1003(t) (97.2 %). strain w3 ...201120584817
a fine control of quorum-sensing communication in agrobacterium tumefaciens.the bacterial pathogen agrobacterium tumefaciens produces the quorum-sensing (qs) signal 3-oxo-octanoylhomoserine lactone (oc8hsl) for controlling horizontal transfer of its tumor inducing (ti) plasmid that carries both the t-dna and the virulence genes. over-accumulation of oc8hsl also increases severity of plant symptoms (number of emerging tumors at infection site) by an unknown mechanism. a. tumefaciens strain c58 expresses two lactonases, attm (blcc) and aiib, that cleave oc8hsl and are pot ...201020585496
pharmacokinetics of opda, an organophosphorus hydrolase, in the african green monkey.organophosphorus (op) pesticides are a broad class of acetylcholinesterase inhibitors that are responsible for tremendous morbidity and mortality worldwide, contributing to an estimated 300,000 deaths annually. current pharmacotherapy for acute op poisoning includes the use of atropine, an oxime, and benzodiazepines. however, even with such therapy, the mortality from these agents are as high as 40%. enzymatic hydrolysis of ops is an attractive new potential therapy for acute op poisoning. a num ...201020599794
activation of an arabidopsis resistance protein is specified by the in planta association of its leucine-rich repeat domain with the cognate oomycete effector.activation of plant immunity relies on recognition of pathogen effectors by several classes of plant resistance proteins. to discover the underlying molecular mechanisms of effector recognition by the arabidopsis thaliana recognition of peronospora parasitica1 (rpp1) resistance protein, we adopted an agrobacterium tumefaciens-mediated transient protein expression system in tobacco (nicotiana tabacum), which allowed us to perform coimmunoprecipitation experiments and mutational analyses. herein, ...201020601497
analysis of core genes supports the reclassification of strains agrobacterium radiobacter k84 and agrobacterium tumefaciens ake10 into the species rhizobium rhizogenes.some strains of the former genus agrobacterium have high biotechnological interest and are currently misclassified. consequently, in this study, the taxonomic status of the non-pathogenic strain agrobacterium radiobacter k84, used in biological control, and the tumourigenic strain agrobacterium tumefaciens ake10, able to regenerate tobacco transgenic plants, was revised. the phylogenetic analysis of the chromosomal genes rrs, atpd and reca showed that they should be reclassified into rhizobium r ...201020627641
a conserved mechanism of gaba binding and antagonism is revealed by structure-function analysis of the periplasmic binding protein atu2422 in agrobacterium tumefaciens.bacterial periplasmic binding proteins (pbps) and eukaryotic pbp-like domains (also called as venus flytrap modules) of g-protein-coupled receptors are involved in extracellular gaba perception. we investigated the structural and functional basis of ligand specificity of the pbp atu2422, which is implicated in virulence and transport of gaba in the plant pathogen agrobacterium tumefaciens. five high-resolution x-ray structures of atu2422 liganded to gaba, pro, ala, and val and of point mutant at ...201020630861
effect of codon optimization and subcellular targeting on toxoplasma gondii antigen sag1 expression in tobacco leaves to use in subcutaneous and oral immunization in mice.codon optimization and subcellular targeting were studied with the aim to increase the expression levels of the sag178-322 antigen of toxoplasma gondii in tobacco leaves. the expression of the tobacco-optimized and native versions of the sag1 gene was explored by transient expression from the agrobacterium tumefaciens binary expression vector, which allows targeting the recombinant protein to the endoplasmic reticulum (er) and the apoplast. finally, mice were subcutaneously and orally immunized ...201020633272
acc deaminase increases the agrobacterium tumefaciens-mediated transformation frequency of commercial canola cultivars.the plant hormone ethylene has been reported to inhibit the agrobacterium tumefaciens-mediated transformation efficiency of many plants. in this study, an acds gene that encodes 1-aminocyclopropane-1-carboxylic acid (acc) deaminase, an enzyme that breaks down acc, the direct precursor of ethylene biosynthesis in all higher plants, was introduced into a. tumefaciens gv3101::pmp90. it was found that the presence of active acc deaminase in a. tumefaciens reduced ethylene levels produced by plant ti ...201020636976
o-methyltransferase(s)-suppressed plants produce lower amounts of phenolic vir inducers and are less susceptible to agrobacterium tumefaciens infection.the first step of agrobacterium tumefaciens/plant interaction corresponds to the activation of a transduction pathway of the bacterium by plant exudate. phenolic compounds rapidly secreted by wounded plant cells induce the expression of bacterial virulence (vir) genes; however, little is known about their biosynthesis in plant. here we show that inoculation of an agrobacterium tumefaciens virulent strain on orthodiphenol-o-methyltransferases-suppressed tobacco plants leads to significantly small ...201020652308
phosphatidylcholine biosynthesis and its significance in bacteria interacting with eukaryotic cells.phosphatidylcholine (pc), a typical eukaryotic membrane phospholipid, is present in only about 10% of all bacterial species, in particular in bacteria interacting with eukaryotes. a number of studies revealed that pc plays a fundamental role in symbiotic and pathogenic microbe-host interactions. agrobacterium tumefaciens mutants lacking pc are unable to elicit plant tumors. the human pathogens brucella abortus and legionella pneumophila require pc for full virulence. the plant symbionts bradyrhi ...201020656373
homologs of aminoacyl-trna synthetases acylate carrier proteins and provide a link between ribosomal and nonribosomal peptide synthesis.aminoacyl-trna synthetases (aarss) are ancient and evolutionary conserved enzymes catalyzing the formation of aminoacyl-trnas, that are used as substrates for ribosomal protein biosynthesis. in addition to full length aars genes, genomes of many organisms are sprinkled with truncated genes encoding single-domain aars-like proteins, which often have relinquished their canonical role in genetic code translation. we have identified the genes for putative seryl-trna synthetase homologs widespread in ...201020663952
bathy phytochromes in rhizobial soil bacteria.phytochromes are biliprotein photoreceptors that are found in plants, bacteria, and fungi. prototypical phytochromes have a pr ground state that absorbs in the red spectral range and is converted by light into the pfr form, which absorbs longer-wavelength, far-red light. recently, some bacterial phytochromes have been described that undergo dark conversion of pr to pfr and thus have a pfr ground state. we show here that such so-called bathy phytochromes are widely distributed among bacteria that ...201020675484
urate is a ligand for the transcriptional regulator pecs.pecs is a member of the marr (multiple antibiotic resistance regulator) family, which has been shown in erwinia to regulate the expression of virulence genes. marr homologs typically bind a small molecule ligand, resulting in attenuated dna binding. for pecs, the natural ligand has not been identified. we have previously shown that urate is a ligand for the deinococcus radiodurans-encoded marr homolog hucr (hypothetical uricase regulator) and identified residues responsible for ligand binding. w ...201020678501
[transformation of enhanced green fluorescent protein gene in paecilomyces lilacinus mediated by agrobacterium tumefaciens].the main aim of this study was to transform the enhanced green fluorescent protein gene (egfp) into biocontrol fungus paecilomyces lilacinus strain 9410. we constructed the expression vector pupngt of the fusion gene nptii-egfp using pcdna3.1(-) as a helper plasmid. the egfp gene was then transformed into p. lilacinus strain 9410 via agrobacterium tumefaciens-mediated transformation. pcr and southern blotting analysis showed that the egfp gene was integrated into the genomes of the tested transf ...201020684307
cholera toxin b subunit-domain iii of dengue virus envelope glycoprotein e fusion protein production in transgenic plants.envelope glycoprotein e of the dengue virus, which plays a crucial role in its entry into host cells, has an immunogenic domain iii (eiii, amino acids 297-394), which is capable of inducing neutralizing antibodies. however, mice immunized with eiii protein without adjuvant elicited low immune responses. to improve low immune responses, a dna fragment, consisting of cholera toxin b subunit and eiii gene (ctb-eiii), was constructed and introduced into tobacco plant cells (nicotiana tabacum l. cv. ...201020691270
agrobacterium-produced and exogenous cytokinin-modulated agrobacterium-mediated plant transformation.agrobacterium tumefaciens is a plant pathogenic bacterium that causes neoplastic growths, called 'crown gall', via the transfer and integration of transferred dna (t-dna) from the bacterium into the plant genome. we characterized an acetosyringone (as)-induced tumour-inducing (ti) plasmid gene, tzs (trans-zeatin synthesizing), that is responsible for the synthesis of the plant hormone cytokinin in nopaline-type a. tumefaciens strains. the loss of tzs protein expression and trans-zeatin secretion ...201020696005
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