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1.
Giorcelli A Sparvoli F Mattivi F Tava A Balestrazzi A Vrhovsek U Calligari P Bollini R Confalonieri M 《Transgenic research》2004,13(3):203-214
When present, stilbene synthase leads to the production of resveratrol compounds, which are major components of the phytoalexin response against fungal pathogens of the plant and are highly bioactive substances of pharmaceutical interest. White poplar (Populus alba L.) was transformed with a construct containing a cDNA insert encoding stilbene synthase from grapevine (Vitis vinifera L.), under the control of the cauliflower mosaic virus (CaMV) 35S promoter, and a chimeric kanamycin resistance gene. Southern blot hybridization analysis demonstrated the presence and integration of exogenous DNA sequences in the poplar genome. Expression of the stilbene synthase-encoding gene in different transgenic lines was confirmed by Western blot and Northern analyses. Compared to the controls, in the transgenic plants two new compounds were detected and were identified as the trans- and cis-isomers of resveratrol-3-glucoside (piceid) by high-pressure liquid chromatography (HPLC), UV spectrophotometry, electrospray mass spectrometry (HPLC-ESI-MS) and enzymatic hydrolysis. Since poplar is a good biomass producer and piceids are accumulated in substantial amounts (up to 615.2 microg/g leaf fresh weight), the transgenic plants represent a potential alternative source for the production of these compounds with high pharmacological value. Despite the presence of piceid, in our experimental conditions no increased resistance against the pathogen Melampsora pulcherrima, which causes rust disease, was observed when in vitro bioassays were performed. 相似文献
2.
Calligari PA Salgado GF Pelupessy P Lopes P Ouazzani J Bodenhausen G Abergel D 《Proteins》2012,80(4):1196-1210
Nuclear magnetic resonance is used to investigate the backbone dynamics in 6-phosphogluconolactonase from Trypanosoma brucei (Tb6PGL) with (holo-) and without (apo-) 6-phosphogluconic acid as ligand. Relaxation data were analyzed using the model-free approach and reduced spectral density mapping. Comparison of predictions, based on 77 ns molecular dynamics simulations, with the observed relaxation rates gives insight into dynamical properties of the protein and their alteration on ligand binding. Data indicate dynamics changes in the vicinity of the binding site. More interesting is the presence of perturbations located in remote regions of this well-structured globular protein in which no large-amplitude motions are involved. This suggests that delocalized changes in dynamics that occur upon binding could be a general feature of protein-target interactions. 相似文献
3.
Delledonne Massimo Allegro Gianni Belenghi Beatrice Balestrazzi Alma Picco Franco Levine Alex Zelasco Samantha Calligari Paolo Confalonieri Massimo 《Molecular breeding : new strategies in plant improvement》2001,7(1):35-42
Transgenic white poplar (Populus alba L.) plants expressing a novel Arabidopsis thaliana cysteine proteinase inhibitor (Atcys) gene have been produced using Agrobacterium tumefaciens-mediated gene transfer. Internodal stem segments of cv. Villafranca were co-cultivated with the EHA105 pBI-Atcys A. tumefaciens strain. Sixteen putative transgenic plant lines were regenerated from different calli with a transformation efficiency of 11%. The integration and expression of the cysteine proteinase inhibitor (Atcys) gene into the plant genome was confirmed by Southern and northern blot analyses. Papain inhibitory activity was detected in poplar transgenic tissues by means of a specific in vitro assay. Such activity was sufficient to inhibit most of the digestive proteinase activity of chrysomelid beetle (Chrysomela populi L.) and confer resistance to C. populi larvae on selected transgenic plants. A close correspondence between the inhibition of papain and resistance to poplar leaf beetle was observed in all tested transgenic lines. Our results indicate that Atcys could be succesfully employed in breeding programmes aimed at the selection of new poplar genotypes resistant to major insect pests. 相似文献
4.
5.
Paolo A. Calligari Gerald R. Kneller Andrea Giansanti Paolo Ascenzi Alessandro Porrello Alessio Bocedi 《Biophysical chemistry》2009
The viral surface glycoprotein neuraminidase (NA) allows the influenza virus penetration and the egress of virions. NAs are classified as A, B, and C. Type-A NAs from influenza virus are subdivided into two phylogenetically distinct families, group-1 and group-2. NA inhibition by oseltamivir represents a therapeutic approach against the avian influenza virus H5N1. Here, structural bases for oseltamivir recognition by group-1 NA1, NA8 and group-2 NA9 are highlighted by the ScrewFit algorithm for quantitative structure comparison. Oseltamivir binding to NA1 and NA8 affects the geometry of Glu119 and of regions Arg130-Ser160, Val240-Gly260, and Asp330-Glu382, leading to multiple NA conformations. Additionally, although NA1 and NA9 share almost the same oseltamivir-bound final conformation, they show some relevant differences as suggested by the ScrewFit algorithm. These results indicate that the design of new NA inhibitors should take into account these family-specific effects induced on the whole structure of NAs. 相似文献
6.
Alma Balestrazzi Silvia Botti Samantha Zelasco Stefania Biondi Cinzia Franchin Paolo Calligari Milvia Racchi Adelaide Turchi Guido Lingua Graziella Berta Daniela Carbonera 《Plant cell reports》2009,28(8):1179-1192
Marker-free transgenic white poplar (Populus alba L., cv ‘Villafranca’) plants, expressing the PsMT
A1
gene from Pisum sativum for a metallothionein-like protein, were produced by Agrobacterium tumefaciens-mediated transformation. The 35SCaMV-PsMT
A1
-NosT cassette was inserted into the ipt-type vector pMAT22. The occurrence of the abnormal ipt-shooty phenotype allowed the visual selection of transformants, while the yeast site-specific recombination R/RS system was
responsible for the excision of the undesired vector sequences with the consequent recovery of normal marker-free transgenic
plants. Molecular analyses confirmed the presence of the 35SCaMV-PsMT
A1
-NosT cassette and transgene expression. Five selected lines were further characterized, revealing the ability to withstand
heavy metal toxicity. They survived 0.1 mM CuCl2, a concentration which strongly affected the nontransgenic plants. Moreover, root development was only slightly affected
by the ectopic expression of the transgene. Reactive oxygen species were accumulated to a lower extent in leaf tissues of
multi-auto-transformation (MAT)-PsMTA1 plants exposed to copper and zinc, compared to control plants. Tolerance to photo-oxidative stress induced by paraquat was
another distinctive feature of the MAT-PsMTA1 lines. Finally, low levels of DNA damage were detected by quantifying the amounts of 8-hydroxy-2′-deoxyguanosine in leaf
tissues of the transgenic plants exposed to copper. 相似文献
7.
Maya M. Polovitskaya Carlo Barbini Diego Martinelli Frederike L. Harms F. Sessions Cole Paolo Calligari Gianfranco Bocchinfuso Lorenzo Stella Andrea Ciolfi Marcello Niceta Teresa Rizza Marwan Shinawi Kathleen Sisco Jessika Johannsen Jonas Denecke Rosalba Carrozzo Daniel J. Wegner Kerstin Kutsche Marco Tartaglia Thomas J. Jentsch 《American journal of human genetics》2020,107(6):1062
8.
Samanta Zelasco Valentina Ressegotti Massimo Confalonieri Daniela Carbonera Paolo Calligari Martina Bonadei Stefano Bisoffi Keiko Yamada Alma Balestrazzi 《Plant Cell, Tissue and Organ Culture》2007,91(1):61-72
Genetic transformation of an elite white poplar genotype (Populus alba L., cv. ‘Villafranca’) was performed with MAT vectors carrying the ipt and rol genes from Agrobacterium spp. as morphological markers. The effects associated with the use of different gene promoters and distinct in vitro regeneration
protocols were evaluated. Poplar plantlets showing abnormal ipt and rol phenotypes were produced only in the presence of exogenous growth regulators. The occurrence of abnormal ipt and rol phenotypes allowed the visual selection of transformants. The ipt-type MAT vector pEXM2 was used to monitor the activity of the yeast site-specific recombination R/RS system in the transformed
white poplar cells. Results from these experiments demonstrated that recombinase-mediated excision events occurred during
the early stages of in vitro culture, thus causing the direct production of ipt marker-free transgenic plants with normal phenotype at an estimated frequency of 36.4%. Beside this unexpected finding, transgenic
ipt-shooty plants were obtained at a frequency of 63.6% and normal shoots were subsequently recovered after a prolonged period
of in vitro culture. Although the transformation efficiency observed in this study, using both ipt and nptII genes as selection markers, was similar to that previously reported with standard vectors carrying only the nptII gene, the easy identification of ipt transformants, the early recombinase-mediated excision events and finally the relatively short time period required to produce
ipt marker-free transgenic plants support for the choice of MAT vectors as a reliable strategy for the future production of marker-free
GM poplars. 相似文献
9.