Search results for "Gel electrophoresi"
showing 10 items of 321 documents
Sequential variation in genomic RNA patterns of human rotaviruses isolated from infantile gastroenteritis
1986
The incidence and RNA electrophoretypes of rotaviral isolates obtained from infants and young children with acute gastroenteritis were studied from October, 1985 through April, 1986. Analysis of the viral RNA was carried out by Polyacrylamide gel electrophoresis followed by silver staining. Fourteen electrophoretypes were identified. A single dominant electrophoretype was found during the first months of the rotavirus seasonal outbreak. In contrast, a large variety of RNA patterns were observed during the latter portion of the study period. Subgrouping of rotavirus isolates by a double-sandwich enzyme-linked immunosorbent assay using monoclonal-detecting antibodies showed that all strains b…
Glycogen phosphorylase from flight muscle of the hawk moth, Manduca sexta: purification and properties of three interconvertible forms and the effect…
1994
Glycogen phosphorylase (EC 2.4.1.1) of Manduca sexta flight muscle was separated into three distinct peaks of activity on diethylaminoethyl-Sephacel. The three fractions of phosphorylase activity were further purified by affinity chromatography on AMP-Sepharose and shown to have the same relative molecular mass (=178000) on polyacrylamide gradient gel electrophoresis under non-denaturating conditions and to produce subunits of molecular mass =92000 on SDS gelelectrophoresis. On the basis of their kinetic properties with respect to the activator AMP and the inhibitor caffeine, the three fractions of phosphorylase activity were assigned as follows: peak 1=phosphorylase b (unphosphorylated for…
Isolation and partial characterization of a cytochrome-o complex from chromatophores of the photosynthetic bacterium Rhodospirillum rubrum FR1.
1989
A cytochrome-o complex was isolated from chromatophores of photoheterotrophically grown Rhodospirillum rubrum FR1. The enzyme was extracted with the non-denaturating detergent taurodeoxycholate and subsequently purified by sucrose-density-gradient centrifugation and gel-permeation HPLC. The complex contains two types of cytochromes, one of them cytochrome o, and two copper atoms. It catalyzes the reduction of molecular oxygen, when N,N,N',N'-tetramethyl-p-phenylenediamine or ubiquinol 10 are offered as electron donors. The oxidase activity is inhibited by cyanide, carbon monoxide and 2-heptyl-2-hydroxyquinoline N-oxide. The molecular mass of the protein is 136 +/- 15 kDa. The subunit analys…
Identification of SCP2165, a new SCP2-derived plasmid of Streptomyces coelicolor A3(2).
2005
Aims: Characterization of SCP2165, a plasmid identified in the Gram-positive bacterium Streptomyces coelicolor A3(2). Methods and Results: Pulsed-field gel electrophoresis (PFGE) of mycelia of a S. coelicolor strain embedded in low melting agarose revealed the presence of a plasmid. Restriction enzyme mapping and sequence analysis of a 2·1 kb fragment revealed that this plasmid could be SCP2. SCP2 and its spontaneous derivative SCP2* are self-transmissible plasmids and have chromosome mobilizing ability (c.m.a.). SCP2* has a c. 1000-fold increased c.m.a. compared with SCP2. Interestingly the plasmid, named SCP2165, shows a c.m.a. from 5 × 10−2 to 1 × 10−1 which is 50–100-fold higher than …
Novel polyaminoacidic copolymers as nonviral gene vectors
2000
Human gene therapy is one of the most promising methods developed in recent years, providing great potential for the treatment of a variety of diseases. Complexes formed between DNA and cationic polymers are attracting increasing attention as novel synthetic vectors for the delivery of genes. We have synthesized polycations with quaternary ammonium groups in their side chains for self-assembly with calf thymus DNA. This paper describes the functionalization of α,β-polyasparthydrazide (PAHy), a synthetic macromolecule having many potential applications in the field of biomedical sciences, with glycidyltrimethylammonuim chloride (GTA) in order to introduce positive charges into their chains. …
Histone variants from pea (Pisum sativum): Their differential presence in fractions obtained by DNase I digestion of nuclei
1990
The variants of the core histones of Pisum sativum L. cv. Lincoln have been resolved by two dimensional polyacrylamide gel electrophoresis. Acetic acid, 8 M urea, 7.2 mM Triton X-100 was used in the first dimension. The second dimension was run in the presence of either anionic (sodium dodecylsulphate) or cationic (cetyltrimethyl-aminonium bromide) detergents. Four putative variants were found for the H2B histone class, 4 for H3 and 3 for H2A. Peptide mapping with (Staphylococcus aureus V8 protease was used, together with other criteria, to characterize the variants. The pattern of histone variants is not organ specific and, in an attempt to determine whether the diversity of histone varian…
Determination of phosphorylation degrees in caseins by on-line gel electrophoresis coupled to ICP-SFMS
2007
The determination of the phosphorylation degree of proteins is essential in many biochemical fields. Besides mass spectrometric techniques elucidating the protein structure, ICP-MS has entered this research area as it provides excellent quantification capabilities. In this work the separation of the proteins has been carried out by gel electrophoresis on-line coupled to an ICP equipped with a double-focusing sector field-MS (ELEMENT 2). Based on the simultaneous detection of 32S+ and 31P+, this work presents an alternative approach to determining the phosphorylation degree of caseins. For the example of α- and β-casein the results obtained were in excellent agreement with the expected value…
Size characterisation of Au nanoparticles by ICP-MS coupling techniques
2006
In modern nanotechnology the properties of nanoparticles (NPs) in different application fields is mainly dependent on their size and their size distribution. This paper aims to develop a reliable method for the size characterisation of Au NPs. Basing on the combination of two different separation techniques (liquid chromatography and gel electrophoresis), coupled on-line to inductively coupled plasma-mass spectrometry (ICP-MS), we study the separation of Au NP standards. Separations by liquid chromatography show size dependent retention behaviour with good reproducibility (RSD < 1%). The results are compared with complementary methods like dynamic light scattering (DLS) and transmission ele…
Determination of molecular mass, Stokes' radius, frictional coefficient and isomer-type of non-denatured proteins by time-dependent pore gradient gel…
1988
Molecular mass, Stoke's radius, frictional coefficient and isomer-type of non-denatured proteins can be obtained by time-dependent gradient gel electrophoresis by evaluating the resulting data using a two-step mathematical procedure. Provided a histochemical staining procedure is available to locate the position of an enzyme in the gel, crude cell extracts can be used for estimating their molecular size properties. The computation of molecular properties of non-denatured proteins is demonstrated for isozymes of aspartate aminotransferase (EC 2.6.1.1), peroxidase (EC 1.11.1.42) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) from current-year needles of spruce. The resulting data as well…
Allozyme electrophoresis: its application to rotifers
1998
Allozyme electrophoresis is a well established technique for revealing genetic variation that has given useful results in a wide range of organisms. However, in rotifers it has been applied scarcely and only in a few species. In this paper the methods of acetate allozyme electrophoresis are introduced, including laboratory setup, equipment and staining recipes that have been successfully applied to brachionid rotifers. In addition, the literature published on allozyme electrophoresis in rotifers is reviewed and the main results and future prospects of the technique are discussed. We conclude that, despite the onset of DNA techniques, allozyme electrophoresis is promising and can yield impor…