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141.
Beaujean A Ducrocq-Assaf C Sangwan RS Lilius G Bülow L Sangwan-Norreel BS 《Biotechnology and bioengineering》2000,70(1):9-16
Manipulation of starch biosynthesis/degradation and formation of novel molecules in storage organs of plants through genetic engineering is an attractive but technically challenging goal. We report here, for the first time, that starch was degraded and glucose and fructose were produced directly when crushed potato tubers expressing a starch degrading bifunctional gene were heated for 45 minutes at 65 degrees C. To achieve this, we have constructed a fusion gene encoding the thermostable enzymes: alpha-amylase (Bacillus stearothermophilus) and glucose isomerase (Thermus thermophilus). The chimeric gene was placed under the control of the granule-bound-starch synthase promoter. This enzymatic complex produced in transgenic tubers was only active at high temperature (65 degrees C). More than 100 independent transgenic potato plants were regenerated. Molecular analyses confirmed the stable integration of the chimeric gene into the potato genome. The biochemical analyses performed on young and old tubers after high-temperature treatment (65 degrees C) revealed an increase in the formation rate of fructose and glucose by a factor of 16.4 and 5. 7, respectively, in the transgenic tubers as compared to untransformed control tubers. No adverse discernible effect on plant development and metabolism including tuber formation and starch accumulation was observed in the transgenic plants before heat treatment. Our results demonstrate that it is possible to replace starch degradation using microbial enzymes via a system where the enzymes are produced directly in the plants, but active only at high temperature, thus offering novel and viable strategies for starch-processing industries. 相似文献
142.
A self-excited biped walking mechanism consisting of two legs that are connected in series at the hip joint through a servomotor is studied as a cyclic system with collisions. A torque proportional to angle between the shank of the swinging leg and the vertical is seen to sustain a gait. Each leg has a thigh and a shank connected at a passive knee joint that has a knee stopper restricting hyperextension similar to the human knee. A mathematical model for the dynamics of the system including the impact equations is used to analyse the stability of the system through examination of phase plane plots. Attractor lines along which the system approaches stability have been identified. A leg length for optimal stability has been identified. The biological basis for the proposed system has been identified by comparison with human gait. 相似文献
143.
144.
Glutamine synthetase in the phloem plays a major role in controlling proline production 总被引:16,自引:1,他引:15 下载免费PDF全文
N Brugiere F Dubois AM Limami M Lelandais Y Roux RS Sangwan B Hirel 《The Plant cell》1999,11(10):1995-2012
To inhibit expression specifically in the phloem, a 274-bp fragment of a cDNA (Gln1-5) encoding cytosolic glutamine synthetase (GS1) from tobacco was placed in the antisense orientation downstream of the cytosolic Cu/Zn superoxide dismutase promoter of Nicotiana plumbaginifolia. After Agrobacterium-mediated transformation, two transgenic N. tabacum lines exhibiting reduced levels of GS1 mRNA and GS activity in midribs, stems, and roots were obtained. Immunogold labeling experiments allowed us to verify that the GS protein content was markedly decreased in the phloem companion cells of transformed plants. Moreover, a general decrease in proline content in the transgenic plants in comparison with wild-type tobacco was observed when plants were forced to assimilate large amounts of ammonium. In contrast, no major changes in the concentration of amino acids used for nitrogen transport were apparent. A (15)NH(4)(+)-labeling kinetic over a 48-hr period confirmed that in leaves of transgenic plants, the decrease in proline production was directly related to glutamine availability. After 2 weeks of salt treatment, the transgenic plants had a pronounced stress phenotype, consisting of wilting and bleaching in the older leaves. We conclude that GS in the phloem plays a major role in regulating proline production consistent with the function of proline as a nitrogen source and as a key metabolite synthesized in response to water stress. 相似文献
145.
Isolation of genomic DNA from defatted oil seed residue of opium poppy (Papaver sominiferum) 总被引:1,自引:0,他引:1
A simple protocol for obtaining pure, restrictable and amplifiable megabase genomic DNA from oil free seed residue ofPapaver sominiferum, an important oil seed and morphanian alkaloid yielding plant, has been developed. Fixed oil from the dry seeds was completely
recovered in an organic solvent and quantified gravimetrically followed by processing of the residual biomass (defatted seed
residue) for genomic DNA isolation. The isolated DNA could be cut by a range of restriction enzymes. The method enables simultaneous
isolation and recovery of lipids and genomic DNA from the same test sample, thus allowing two independent analyses from a
single sample. 相似文献
146.
Certain drugs and chemicals, such as chloroquine, chlorpromazine, and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), are bound to melanin and retained in pigment cells for long periods. This specific retention in pigmented tissues can cause adverse effects in the skin, eye, inner ear, and pigmented nerve cells of the substantia nigra of the brain. To date, all studies have been focused on eu- and neuromelanin. In the present study, we show that chloroquine, chlorpromazine, chlomipramine, paraquat, acridine orange, and nickel, which are bound to eumelanin, also bind to synthetic pheomelanin, but the binding to pheomelanin is lower. The binding varied with the cysteine content and pH, and the results indicate that the binding is complex and includes ionic interactions. In addition, we have shown that these substances also bind to synthetic thiourea-containing melanin, but to quite a low extent. We also present a microautoradiographic study on the binding of 14C-chloroquine to natural pheomelanin in vivo in yellow mice C57BL (Ay/a). Black (C57/BL) and albino (NMRI) mice were used as controls. The autoradiography demonstrated a pronounced uptake of chloroquine in the hair follicles and the dermal melanocytes in the ear of yellow mice, which was comparable to the corresponding accumulation of label in black mice. In the albino mouse, the uptake was lower and more homogeneously distributed in the skin. These results suggest that the toxicologi-cal risks of melanin-related adverse effects are applicable to persons with a high content of pheomelanin in the skin and hair. 相似文献
147.
148.
SV40 large T antigen has been reported to be modified with several
different sugars including N-acetylglucosamine, galactose, and mannose. In
this report we have reexamined the glycosylation of T antigen and found
that while we could detect modification with N-acetylglucosamine, we could
not detect any other sugars on the protein. Surprisingly, even though
[3H]galactose could be metabolically incorporated into the protein,
analysis showed that all of the radioactivity in T antigen had been
converted to other species. The N-acetylglucosamine was demonstrated to be
linked to the protein in the form of O-linked N- acetylglucosamine, the
best characterized form of nuclear and cytoplasmic glycosylation in
mammalian systems. We have localized the major site of glycosylation to the
amino terminal portion of the molecule. Analysis of mutated T antigen where
serines 111/112 were substituted with alanine suggest that these residues
constitute a glycosylation site on the protein. These two serines fall
within a typical O-linked N-acetylglucosamine glycosylation site (PSS) and
are also known to be phosphorylated. Thus, it is likely that competition
between phosphorylation and glycosylation occurs at this site.
相似文献
149.
150.
Ghosh Mayukh Sangwan N. Chakravarti S. Banerjee Somesh Ghosh A. Kumar R. Sangwan A. K. 《International journal of peptide research and therapeutics》2019,25(4):1291-1308
International Journal of Peptide Research and Therapeutics - Vector-borne diseases, mainly transmitted by ticks, have a significant impact on the productivity and health of the animals in the... 相似文献