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排序方式: 共有206条查询结果,搜索用时 15 毫秒
1.
Jan Schripsema Annemarie H. Meijer Frank van Iren Hens J. G. ten Hoopen Robert Verpoorte 《Plant Cell, Tissue and Organ Culture》1990,22(1):55-64
A simple non-invasive method for the characterization of growth of a plant cell suspension in a single culture flask is given. The dissimilation of sugars by a cell-culture causes a loss of weight of the contents of the culture flask, and can therefore be used to follow the growth in that single culture flask. Because a correction for water evaporation is necessary, accurate results can only be obtained when a stable closure is used (e.g. Silicosen T-type plugs). The dissimilation curves obtained in this way were correlated to the concentration of sugars in the medium, the dry weight and the fresh weight. From these correlations the amount of intracellularly stored carbohydrates could be estimated. Rate constants for CO2-diffusion were determined for different types of closure. These values allowed the estimation of CO2 levels inside the culture flasks from the dissimilation curves (CO2 release curves). The dissimilation curves obtained using this method can easily be related to other types of growth curves. Different growth-phases can be clearly distinguished, e.g. lag-phase, exponential growth-phase and stationary-phase. 相似文献
2.
Charlotte Poulsen Oscar J. M. Goddijn J. Harry C. Hoge Robert Verpoorte 《Transgenic research》1994,3(1):43-49
TransgenicNicotiana tabacum L. Petit Havana SR1 F1-plants expressing tryptophan decarboxylase cDNA (tdc) fromCatharanthus roseus (L.) G. Don under the control of the CaMV 35S promoter and terminator exhibited tryptophan decarboxylase (TDC) enzyme activity and accumulated tryptamine. The plants with the highest TDC activity contained 19 pkat per mg of protein. The influence of transgenic expression oftdc on the activities of anthranilate synthase (AS) and chorismate mutase (CM) were examined in 10 transgenic tobacco plants. The specific activities of these two chorismate-utilizing enzymes were not significantly affected by expression oftdc, despite their important functions as branch point enzymes in the shikimate pathway. The results indicate that the normal route of tryptophan biosynthesis in plants is sufficient to supply a considerable amount of this essential amino acid for the biosynthesis of secondary metabolites. Despite their increased tryptamine content, the growth and development of the transgenic tobacco plants expressingtdc appeared normal. 相似文献
3.
4.
Dhavadee Ponglux Tanomjit Supavita Robert Verpoorte David Phillipson 《Phytochemistry》1980,19(9):2013-2016
The major alkaloids of a sample of leaves of Uncaria attenuata obtained from Thailand have been identified as the pentacyclic heteroyohimbine alkaloids tetrahydroalstonine, rauniticine and the novel 14-β-hydroxy-3-iso-rauniticine. Evidence for the structure of the new alkaloid was obtained from a study of UV, IR, MS, 1H NMR and 1C NMR spectra. 相似文献
5.
Bacterial spinae from marine bacterium D71 are multi-subunit structures of a single protein. This protein, called spinin, is homogeneous by immunodiffusion and immunoelectrophoresis, amino acid composition, polyacrylamide gel electrophoresis with a number of buffer systems, sedimentation velocity and diffusion boundary analysis. Sedimentation equilibrium gives Mr = 19,000, while phosphate polyacryl-amide gel electrophoresis in presence of dodecyl sulfate gives Mr = 32,000. The lower Mr estimate for spinin is supported by sedimentation equilibrium in 6 M guanidine . HCl, and covalent cross-linking with dimethyl suberimidate or glutaraldehyde. The higher Mr value probably arises from an anomalous spinin-dodecyl sulfate interaction. Isoelectric focusing in polyacrylamide gel gives pI = 3.45; however, the focusing pattern also contains three distinct bands that may arise from hydrolysis of the spinin protomer during anodic migration. This study presents the first extensive physicochemical characterization of spinin and provides the basis for investigating the subunit assembly of spinae. 相似文献
6.
7.
Metabolomics is defined as both the qualitative and quantitative analysis of all metabolites in an organism unraveling correlation
with other OMICs data. Many of the technologies used in metabolomics have method-specific advantages and drawbacks in terms
of diversity of metabolites detected, sensitivity, or resolution. In this paper, the potential of NMR spectrometry applied
to metabolomics is reviewed using examples of Nicotiana tabacum and Catharanthus roseus. 相似文献
8.
Here the current status of knowledge on some well-characterized transporters located in the vacuolar membrane is reviewed.
As different cellular compartments and even different cells may be involved in certain steps of a biosynthetic pathway, the
regulation of the flux is not only dependent on structural genes encoding enzymes catabolizing certain steps but also transport
has a major regulatory function. The aim of the present review is to give an overview of the present knowledge of transport
of secondary metabolites in plants, and to use this information in the context of our knowledge about Catharanthus roseus alkaloid biosynthesis. This should lead to further insight in the possible role of various transporters in the regulation
of the biosynthesis of these alkaloids. 相似文献
9.
A microplate assay for mevalonate and 5-phosphomevalonate kinase activities has been developed using an enzyme-coupled system of pyruvate kinase and lactate dehydrogenase. Mevalonate and 5-phosphomevalonate kinase activities were measured in crude and partially purified enzyme preparations from Catharanthus roseus suspension-cultured plant cells. The assay was validated with respect to protein and substrate concentration. Mevalonate and 5-phosphomevalonate kinase showed Michaelis-Menten kinetics with respect to ATP and their specific substrates; the apparent Km values of mevalonate kinase for ATP and mevalonate were 210 and 65 microM, respectively, and of 5-phosphomevalonate kinase for ATP and 5-phosphomevalonate were 0.41 and 0.4 mM, respectively. Considering mevalonate kinase, the relative standard deviation of enzyme activity within a determination (n = 3) is always less than 2.5% and in between determinations (n = 9) is less than 2%. The method can be used in a continuous assay as well as in a discontinuous assay. 相似文献
10.
Rafael Zárate Johan Memelink Robert van der Heijden Robert Verpoorte 《Biotechnology letters》1999,21(11):997-1002
In vitro propagation of Catharanthus roseus was achieved using nodal explants. Bud induction was best on medium containing 1.0 mg benzyl aminopurine l–1. Hardening of rooted shoots to soil was very successful with 98% survival. Genetically transformed C. roseus plantlets were obtained after bombardment of nodal explants, which were then micropropagated, with DNA coated particles with green fluorescent protein (GFP) or -glucuronidase (GUS) reporter genes. Histological studies showed that the gene insertion method proved effective with many cells and different tissues displaying the reporter gene signals, showing that gene expressions were rather stable. 相似文献