Cultural characterization

The rhizosphere microorganisms can form beneficial, pathogenic, or neutral relationships. These relationships can promote plant growth and productivity. Among them, a number of Variovorax isolates from the rhizosphere were isolated. Bacteria Variovorax strain C6d (AB552893) was isolated from the non axenic culture of Chlorella spp., C6.

8p vibecca 01-10-2024 3 1   Download

Azospirillum is one of the versatile non-symbiotic, free living diazotrophic bacteria which appears to have a world-wide distribution and occurs in large number in the rhizosphere soil of a variety of grasses and cereals. The present study was carried out during Rabi and Kharif 2016-17 in the Department of Soil Science and Agricultural Chemistry, Birsa Agricultural University, Ranchi, Jharkhand. Efforts were made to screen out the presence of Azospirillum in rhizosphere of various non-leguminous crops and to characterize the isolates on the basis of morphological and cultural behaviours.

10p cothumenhmong5 17-05-2020 14 4   Download

Many groups of proteins play important roles in the cell’s response to various stresses. The molecular chaperone GroEL of Escherichia coli represents one such highly conserved family of stress proteins. We have observed that isolated GroEL complexes from stationary cultures contain various polypeptides that can be released from the chaperonin by GroES and/or ATP, and identified two such polypeptides as the proteins GatY and UP12.

9p system191 01-06-2013 28 3   Download

An oxidosqualene cyclase cDNA, LcIMS1, was isolated from cultured cells of Luffa cylindrica Roem. by heterologous hybridization with cDNA of Glycyrrhiza glabra b-amyrin synthase. Expression of LcIMS1 in yeast lacking endogenous oxidosqualene cyclase activity resulted in the accumulation of isomultiflorenol, a triterpene. This is consistent with LcIMS1 encoding isomultiflorenol synthase, an oxidosqualene cyclase involved in bryonolic acid biosynthesis in cultured Luffa cells.

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The octadecaneuropeptide (ODN; QATVGDVNTDRPG LLDLK) and its C-terminal octapeptide (OP; RPGLLDLK), which exert anxiogenic activity, have been previously shown to increase intracellular calcium concentration ([Ca21]i) in cultured rat astrocytes through activation of a metabotropic receptor positively coupled to phospholipase C. It has also been found that the [D -Leu5]OP analog possesses a weak antagonistic activity. The aim of the present study was to synthesize and characterize cyclic analogs of OP and [D -Leu5]OP.

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Two secreted acid phosphatases (SAP1 and SAP2) were markedly up-regulated during Pi-starvation of tomato suspension cells.SAP1 and SAP2 were resolved during cat-ion-exchange FPLC of culture media proteins from 8-day-old Pi -starved cells, and purified to homogeneity and final p-nitrophenylphosphatehydrolyzing specificactivitiesof 246 and 940lmol Pi producedÆmin )1 mgÆprotein )1 , respect-ively.SDS/PAGE, periodic acid-Schiff staining and analyt-ical gel filtration demonstrated that SAP1 and SAP2, respectively, exist as 84 and57 kDa glycosylatedmonomers....

9p tumor12 22-04-2013 54 2   Download

Selenium (Se) is well known to be essential for cell culture when using a serum-free medium, but not when a medium containing serum is used. This finding suggests that serum contains some usable formof Se. To identify the Se-supplier, T-lymphoma (Jurkat) cells were cultured for 3 days in the presence of human serum immunodepleted of Se-containing serum protein, selenoprotein Por extracellular glutathione peroxidase.

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ATP-regenerating enzymes may have an important role in maintaining ATP levels in mitochondria-like kinetoplast organelle and glycosomes in parasitic protozoa. Adenylate kinase (AK) (ATP:AMP phosphotransferase) catalyses the reversible transfer of thec-phosphate group from ATP to AMP, releasing twomolecules ofADP. This study describes cloningand functional characterizationof the gene encoding AK2fromagenomic libraryofLeishmaniadonovaniandalso its expression in leishmania promastigote cultures.

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A high-affinity monoclonal antibody (M27), raised against the human thrombin–antithrombin complex, has been identifiedand characterized. The epitope recognizedbyM27 was located to the linear sequence FIREVP (residues 411– 416), located in the C-terminal cleavage peptide of anti-thrombin.This regionoverlaps, by two residues, theputative binding site of antithrombin for the serpin–enzyme complex receptor.

11p tumor12 20-04-2013 37 4   Download

In this article, we report the results of an analysis of the glycolytic enzyme enolase (2-phospho-D-glycerate hydro-lase) of Trypanosoma brucei. Enolase activity was detected in both bloodstream-form and procyclic insect-stage try-panosomes, although a 4.5-fold lower specific activity was found in the cultured procyclic homogenate. Subcellular localization analysis showed that the enzyme is only pre-sent in the cytosol.

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We have previously cloned, expressedand characterized two variants of the major allergen Lep d 2 from culturedLepi-doglyphus destructor mites. These variants, Lep d 2.0101 and Lep d 2.0201, differ at 13 amino acid positions. In this study we investigated Lep d 2 sequence diversity between wild and cultured mites. PCR, Southern blot and DNA sequence analysis revealed the presence of two different Lep d 2 genes, one with and one without an intron.

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We have isolated and characterized a second [Fe]-hydro-genasegene fromthegreenalga,Chlamydomonas reinhardtii. TheHydA2gene encodes a protein of 505 amino acids that is 74% similar and 68% identical to the known HydA1 hydrogenase fromC. reinhardtii.HydA2 contains all the conserved residues and motifs found in the catalytic core of the family of [Fe]-hydrogenases. We demonstrate that both theHydA1and theHydA2transcripts are expressed upon anaerobic induction, achieved either by neutral gas purging or by sulfur deprivation of the cultures. ...

9p fptmusic 16-04-2013 46 2   Download

A thermostable enzyme from the hyperthermophilic sul-phate-reducing archaeon, Archaeoglobus fulgidus,was expressed and characterized on the assumption that it is homologous to exonuclease III from Escherichia coli. Sequence similaritydatabase searcheswere performedbased on the amino acid sequence of exonuclease III. The 774bp longgenewas isolated froma culture sample andcloned into different vectors.

12p fptmusic 16-04-2013 27 3   Download

For a long time, the haemerythrin family of proteins was considered to be restricted to only a few phyla of marine invertebrates. When analysing dif-ferential protein expression in the methane-oxidizing bacterium, Methylo-coccus capsulatus(Bath), grown at a high and low copper-to-biomass ratio, respectively, we identified a putative prokaryotic haemerythrin expressed in high-copper cultures.

0p awards 06-04-2013 30 1   Download

We have cloned, over-expressed and purified enolase from Plasmodium falciparumstrain NF54 in Escherichia coliin active form, as an N-terminal His6-tagged protein. The sequence of the cloned enolase from the NF54 strain is identical to that of strain 3D7 used in full genome sequen-cing. The recombinant enolase (r-Pfen) could be obtained in large quantities (
50 mg per litre of culture) in a highly purified form ( 95%). The purified protein gave a single band at
50 kDa on SDS/PAGE.

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We describe the expression, purification, and biochemical characterization of two homologous enzymes, with amido-hydrolase activities, of plant (Lupinus luteuspotassium-independent asparaginase, LlA) and bacterial (Escherichia coli, ybiK/spt/iaaA gene product, EcAIII) origin. Both enzymes were expressed inE. colicells, with (LlA) or with-out (EcAIII) aHis-tag sequence. The proteins were purified, yielding6 or 30 mgÆL )1 of culture, respectively.

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GM-CSF is one of several naturally occurring glycoproteins that regulate leukocyte production, migration and function. It has been produced in different cell types, with different properties that depend on the production process used. The purpose of this work was to characterize the recombinant human GM-CSF from an engineered Chinese hamster ovary cell line grown in suspension and as adherent culture for the identification of the glycosylation sites and the defi-nition of the glycosidic moiety, including the degree of site occupancy....

13p dell39 03-04-2013 46 4   Download

Bacillus cereusisolated from the larvae of Myrmeleon bore was found to secrete proteins that paralyze and kill German cockroaches, Blattela germanica, when injected. One of these active proteins was purified from the culture broth ofB.cereususing anion-exchange and gel-filtration chro-matography. The purified toxin, with a molecular mass of 34 kDa, was identified as sphingomyelinase C (EC3.1.4.12) on the basis of its N-terminal and internal amino-acid sequences. A recombinant sphingomyelinase C expressed in Escherichia coliwas as potent as the native protein in killing the cockroaches....

6p dell39 03-04-2013 48 4   Download

Two soluble enzymes (FerA and FerB) catalyzing the reduction of a number of iron(III) complexes by NADH, were purified to near homogeneity from the aerobically grown iron-limited culture ofParacoccus denitrificansusing a combination of anion-exchange chromatography (Seph-arose Q), chromatofocusing (Mono P), and gel permeation chromatography (Superose 12). FerA is a monomer with a molecular mass of 19 kDa, whereas FerB exhibited a molecular mass of about 55 kDa and consists of probably two identical subunits. ...

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We have isolated a laccase (lac1) from culture fluid ofVol-variella volvacea, grown in a defined medium containing 150lMCuSO4 , by ion-exchange and gel filtration chroma-tography. Lac1 has a molecular mass of 58 kDa as deter-mined by SDS/PAGE and an isoelectric point of 3.7. Degenerate primers based on the N-terminal sequence of purified lac1 and a conserved copper-binding domain were used to generate cDNA fragments encoding a portion of the lac1 protein and RACE was used to obtain full-length cDNA clones. ...

11p dell39 03-04-2013 56 6   Download