Isolation and characterization of bioactive compounds from plant resources: The role of analysis in the ethnopharmacological approach

2014 ◽  
Vol 87 ◽  
pp. 218-228 ◽  
Author(s):  
G. Brusotti ◽  
I. Cesari ◽  
A. Dentamaro ◽  
G. Caccialanza ◽  
G. Massolini
Keyword(s):  
Bioactive Compounds ◽  
Plant Resources ◽  
Isolation And Characterization

Isolation and characterization of SRF accessory proteins

1993 ◽  
Vol 340 (1293) ◽  
pp. 325-332 ◽  
Keyword(s):  
Ternary Complex ◽  
Map Kinases ◽  
Serum Response Factor ◽  
Regulatory Element ◽  
Isolation And Characterization ◽  
Activation Of Transcription ◽  
Serum Response ◽  
Dna Binding Properties

Many genes which are regulated by growth factors contain a common regulatory element, the serum response element (SRE). Activation of transcription by the SRE involves a ternary complex formed between a ubiquitous factor, serum response factor (SRF), and a second protein, p62/TCF. We used a yeast genetic screen to isolate cDNAs encoding a protein, SAP-1, with the DNA binding properties of p62/TCF. The SAP-1 sequence contains three regions of homology to the previously uncharacterized Elk-1 protein, which also acts as an SRF accessory protein. Only two of these regions are required for cooperative interactions with SRF in the ternary complex. The third contains several conserved sites for the MAP kinases, whose activity is regulated in response to growth factor stimulation. We discuss the potential role of these proteins in regulation of the c-fos SRE.

The role of Golgi bodies in polysaccharide sulphation in Fucuszygotes

1978 ◽  
Vol 32 (1) ◽  
pp. 337-356
Author(s):  
M.E. Callow ◽  
S.J. Coughlan ◽  
L.V. Evans
Keyword(s):  
Cell Wall ◽  
Alginic Acid ◽  
Soluble Fraction ◽  
Amorphous Layer ◽  
Acceptor Molecule ◽  
Golgi Bodies ◽  
Isolation And Characterization ◽  
Fucus Serratus

The cell wall of 24-h zygotes of Fucus serratus is composed of 3 layers—an inner fibrillar layer (sulphated fucan), an outer fibrillar layer (alginic aicd/cellulose) and an exterior amorphous layer (sulphated fucan, alginic acid). The 2 layers containing sulphated fucan are preferentially thickened at the rhizoid pole. Light- and electron-microscope autoradiographic pulse-chase experiments on 22-h zygotes using 35SO2-(4) show the Golgi bodies to be the sites of fucan sulphation. The isolation and characterization of isolated Golgi-rich fractions from 22-h zygotes shows that the first detectable labelled macromolecule is associated with these fractions 2 min after addition of 35SO2-(4). The sulphate acceptor molecule has been partially characterized. 35S-APS and 35S-paps are detectable in the soluble fraction 0.5 min after addition of 35SO2-(4). The results are discussed in relation to other published work on the differentiation of Fucus embryos and on polysaccharide sulphation.

scholarly journals Isolation and Characterization of the PISTILLATA Ortholog Gene from Cymbidium faberi Rolfe

Agronomy ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 425 ◽  
Author(s):  
Yue Fei ◽  
Zhi-Xiong Liu
Keyword(s):  
Floral Development ◽  
Economic Value ◽  
Ectopic Expression ◽  
Anther Development ◽  
Protein Alignment ◽  
Isolation And Characterization ◽  
Petal Development ◽  
Potted Plant

Cymbidium faberi Rolfe is a very popular potted plant in China, Japan and Korea where it has been cultivated for centuries. The economic value of this popular native Asian orchid could be enhanced by changes in its floral traits. In Arabidopsis, PISTILLATA (PI) is involved in regulating petal and stamen development. In order to investigate the possible role of the PI ortholog involved in floral development, we isolated CyfaPI from C. faberi. Protein alignment and a phylogenetic tree grouped CyfaPI in the PI lineage. CyfaPI transcripts were detected in all floral organs, but were absent in leaves. Moreover, in flowers, the highest expression level of CyfaPI was present in the gynostemium and the lowest level was found in anther caps. In addition, ectopic expression of CyfaPI in Arabidopsis pi-1 mutant rescued petal development, and complement the development of filament-like structure (part of stamen), but failed to complement anther development in the stamen whorl. All these finding suggest that CyfaPI is mainly responsible for perianth and gynostemium development in C. faberi. Our data may help to trace the development of the gynostemium program and evolution in orchids.

Dicoumarol-induced prothrombins containing 6, 7, and 8 γ-carboxyglutamic acid residues: isolation and characterization

1989 ◽  
Vol 67 (8) ◽  
pp. 411-421 ◽  
Author(s):  
Om P. Malhotra
Keyword(s):  
Gel Electrophoresis ◽  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Calcium Dependent ◽  
Isolation And Characterization ◽  
Carboxyglutamic Acid ◽  
K Deficiency ◽  
Agar Gel Electrophoresis

Isolation and characterization of γ-carboxyglutamic acid (Gla) deficient prothrombins induced by Warfarin or dicoumarol are useful for studying the role of specific Gla residues in prothrombin. In addition to 7-Gla prothrombin, we have isolated two more atypical prothrombins from the barium citrate eluate, one containing 6.11, and the other, 7.85 Gla residues, presumably 6- and 8-Gla prothrombins. The actual Gla content of the 7-Gla isomer was 7.05. Each of the 6-, 7-, and 8-Gla variants showed a single component by agar or dodecyl sulfate Polyacrylamide gel electrophoresis. When agar gel electrophoresis was performed in calcium, each of the variants moved more rapidly than normal (10-Gla) prothrombin. In the presence of EDTA, the 8-Gla isomer exhibited the fastest mobility, equivalent to that of normal prothrombin, followed by 7-, and then 6-Gla variants. The physiological activities of the isomers were found to be 18 to 23% for 8-, 6 to 8% for 7-, and 2 to 3% of normal prothrombin for 6-Gla variant. Prothrombin fragment 1, derived from 8-Gla prothrombin, exhibited 23% of calcium-induced fluorescence quenching, compared with 40% for 10-Gla and 8% or less for 7- and 6-Gla fragments 1. Competition radioimmunoassay data show that calcium-dependent anti (normal) prothrombin polyclonal antibodies are not specific for 10-Gla prothrombin, since the 7- and 8-Gla isomers were able to displace radiolabeled (125I) normal prothrombin.Key words: prothrombin, blood clotting, dicoumarol, Warfarin, γ-carboxyglutamic acid, vitamin K deficiency.

scholarly journals Isolation and Characterization of Cryptococcus neoformans Spores Reveal a Critical Role for Capsule Biosynthesis Genes in Spore Biogenesis

2009 ◽  
Vol 8 (4) ◽  
pp. 595-605 ◽  
Author(s):  
Michael R. Botts ◽  
Steven S. Giles ◽  
Marcellene A. Gates ◽  
Thomas R. Kozel ◽  
Christina M. Hull
Keyword(s):  
Cryptococcus Neoformans ◽  
Fungal Pathogens ◽  
Critical Role ◽  
Spore Formation ◽  
Yeast Cells ◽  
Isolation And Characterization ◽  
Specific Configuration ◽  
First Time

ABSTRACT Spores are essential particles for the survival of many organisms, both prokaryotic and eukaryotic. Among the eukaryotes, fungi have developed spores with superior resistance and dispersal properties. For the human fungal pathogens, however, relatively little is known about the role that spores play in dispersal and infection. Here we present the purification and characterization of spores from the environmental fungus Cryptococcus neoformans. For the first time, we purified spores to homogeneity and assessed their morphological, stress resistance, and surface properties. We found that spores are morphologically distinct from yeast cells and are covered with a thick spore coat. Spores are also more resistant to environmental stresses than yeast cells and display a spore-specific configuration of polysaccharides on their surfaces. Surprisingly, we found that the surface of the spore reacts with antibodies to the polysaccharide glucuronoxylomannan, the most abundant component of the polysaccharide capsule required for C. neoformans virulence. We explored the role of capsule polysaccharide in spore development by assessing spore formation in a series of acapsular strains and determined that capsule biosynthesis genes are required for proper sexual development and normal spore formation. Our findings suggest that C. neoformans spores may have an adapted cell surface that facilitates persistence in harsh environments and ultimately allows them to infect mammalian hosts.

scholarly journals Isolation and Characterization of Human Lung Lymphatic Endothelial Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Bruno Lorusso ◽  
Angela Falco ◽  
Denise Madeddu ◽  
Caterina Frati ◽  
Stefano Cavalli ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Tyrosine Kinases ◽  
Human Lung ◽  
Lymphatic Endothelial Cells ◽  
Isolation And Characterization ◽  
Microbial Agents ◽  
Diseased Lung

Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retainin vitroseveral receptor tyrosine kinases (RTKs) implicated in cell survival and proliferation. HL-LECs represent a clinically relevant cellular substrate to study lymphatic biology, lymphoangiogenesis, interaction with microbial agents, wound healing, and anticancer therapy.

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