摘要:

AbstractTo estimate the feasibility of isoelectric focusing in a continuous mode, the conditions for obtaining stable and linear pH gradients were first investigated, using a mixture of carrier ampholytes. Then the operating parameters allowing the isoelectric focusing of bovine serum albumin as model protein were determined. A theoretical approach was used to find conditions minimizing the electrohydrodynamic effect, mainly responsible for protein dispersion, for the separation of bovine serum albumin and α‐lactalbumin from bovine mi

ISSN:0173-0835    

DOI:10.1002/elps.1150190813

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractFor isolation of low soluble recombinant full‐length (amino acids 1–191) core protein of hepatitis C virus (HCV) overexpressed inEscherichia coli, the advantage of combining two electrophoretic techniques, in comparison with chromatographic separation, is demonstrated. The protein extract was first solubilized in agents compatible with electrophoretic separation. Using preparative liquid phase isoelectric focusing (IEF) the protein of interest was first concentrated within a defined acidic pH range. These fractions were then submitted to preparative sodium dodecyl sulfate ‐ polyacrylamide gel electrophoresis (SDS‐PAGE) to isolate the 22 kDa protein. The second‐dimensional step allowed the isolation of 2 mg of the purified recombinant HCV core protein (rHCV‐C191) from 1.5 g bacterial pellet. This quantity is sufficient to characterize the protein and to perform immunogenicity studies. This procedure of two‐dimensional preparative electrophoresis is applicable to a wide range of biological samples and represents an alternative for purification of insol

ISSN:0173-0835    

DOI:10.1002/elps.1150190814

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractCreutzfeldt‐Jakob disease (CJD) is now recognized to occur in four main forms: classical sporadic, genetic, iatrogenic and new variant. These are briefly described. The evidence that new variant disease is due to bovine spongiform encephalopathy (BSE) is reviewed. In CJD, disease and infectivity are associated with the accumulation of an abnormal form of a normal cellular protein. Protein analysis and the study of the gene which encodes for this protein have therefore played an important role in understanding CJD. Diagnostic tests for CJD are discussed with particular reference to the detection of the 14‐3‐3 protein in the cerebrospinal fluid

ISSN:0173-0835    

DOI:10.1002/elps.1150190815

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractThe effects of agarose gel strips without and with 0.03mrelectroendosmosis (EEO) on isoelectric focusing (IEF) were studied. It is shown that only agarose without EEO can be used for IEF. The effects of electrode buffer strips using agarose with different EEO of 0, 0.03, 0.08, 0.20mron sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and native PAGE were also studied. It apparently did not affect SDS‐PAGE, but affected native PAGE to a certain extent. The higher the EEO value was, the more water was present on the agarose gel strip during a separation. Agarose gel strips with an EEO of 0.2mrare not suitable as electrode buffer strips for native

ISSN:0173-0835    

DOI:10.1002/elps.1150190816

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractIsoelectric focusing in immobilized pH gradients ranging from pH 5.05 to 5.60 was used to study the distribution of transferrin (Tf) subtypes and their gene frequencies from 188 unrelated healthy donors of the Han population in Beijing. Six phenotypes (TfCl, TfC2, TfC1C2, TfC1C3, TfC1Dchiand TfC2Dchi) were detected and Tf*C3 has never before been reported in the Han population. Gene frequencies were as follows: Tf*Cl = 0.7420, Tf*C2 = 0.2420, Tf*C3 = 0.0027, Tf*Dchi = 0.0133. There is good agreement between the observed and expected values corresponding to the Hardy‐Weinberg equilibrium (∑χ2= 0.9183,df= 2, 0.5

ISSN:0173-0835    

DOI:10.1002/elps.1150190817

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractComponents of the crude extract of barley, recognized by IgE antibodies in sera from 132 pig farm‐workers in southern Taiwan, were investigated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and immunoblotting. Among the sera tested, 43 (32.6%) showed IgE reactivity to 15 components of barley with molecular mass ranging from 148000 to 14000 daltons (Da). Heterogeneity in the IgE‐binding patterns of the samples tested was observed. However, the major antigen appears to be the component with about 37 kDa, in 32 (74.4%) of the 43 immunoblot‐positive serum samples. Furthermore, components with about 55 and 67 kDa which reacted with 26 (60.5%) and 18 (41.9%) of the positive samples, respectively. These might also be considered as important allergens of the flour made from entire seeds of barley. The results suggest that barley may be considered to play a role in the IgE‐mediated occupational respiratory disorders and the 37 kDa component has also been shown to be the maj

ISSN:0173-0835    

DOI:10.1002/elps.1150190818

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractWe used three different electrophoretic techniques for the analysis of human plasma proteins: (i) two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE), with sodium dodecyl sulfate (SDS) used only in slab gel electrophoresis; (ii) capillary isoelectric focusing (CIEF) with no denaturants; (iii) linear polyacrylamide (LPA)‐filled capillary electrophoresis with SDS (SDS‐CE). With technique (i), data on isoelectric point and molecular size of plasma proteins can be obtained. Techniques (ii) and (iii) are suited to obtain quantitative information on proteins. The separation principle used in technique (ii) is closely related to that used in the first dimension of technique (i), and that used in technique (iii) related to that in the second dimension of technique (i). Therefore, we could successfully correlate protein separation patterns obtained by 2‐D PAGE and those obtained by capillary electrophoresis. The advantages of correlating data obtained by various electrophoretic techniques in the course of constructing a comprehensive database on human plasma proteins are

ISSN:0173-0835    

DOI:10.1002/elps.1150190819

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractNatural resistance toMycobacterium bovisbacillus Calmette‐Guérin (BCG) is determined by theBcggene (Nramp1), which is exclusively expressed by mature macrophages. TheNramp1gene is a dominant autosomal gene that has two allelic forms;rconfers resistance andsconfers susceptibility to infection with intracellular pathogen. Although the wide range of pleiotropic immunological effects of theNramp1gene has been described, the exact mechanism of its action remains elusive. In this study we searched for differentially expressed proteins that might provide clues in the studies onNramp1gene function. We performed two‐dimensional gel electrophoresis of cellular proteins prepared from a B10R macrophage line derived from mice carrying therallele of theNramp1gene, B10S macrophages carrying thesallele, and B10R‐Rb macrophages transfected withNrampl‐ribozyme. The classification of protein patterns and selection of distinct proteins characteristic ofrorsallele‐carrying macrophages was performed using the principal component analysis. We found differential expression of four proteins with the following isoelectric point/molecular weight (pI/Mr) in B10R macrophages compared to B10S and B10R‐Rb macrophages: 6.6/25, 7.0/22, 9.1/31.5, and 5.3/8.5. The protein 7.0/22 has been identified as Mn‐superoxide dismutase and the best candidate for protein p6.6/25 seems to be Bcl‐2 according to the immunoblot analysis. When the splenic macrophages carring therorsallele were analyzed, the changes in relative abundance for proteins 6.6/25 and p7.0/22 were satisfactorily reproduced. Overall, the two identified proteins are important in the regulation of intracellular redox balance and the regulation of apoptosis in macrophages, respectively. Our findings may suggest their possible biological role in the innate immunity against intrace

ISSN:0173-0835    

DOI:10.1002/elps.1150190820

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractGlutathioneS‐transferases (GST) are a family of detoxification isoenzymes that catalyze the conjugation of xenobiotics and their metabolites with reduced glutathione. Lead exposure in rats is known to induce GST isoenzymes in the liver and kidney. These changes in expression have potential use as biomarkers of lead exposure. Because two‐dimensional electrophoresis (2‐DE) enables one to analyze both protein abundance changes and chemical changes in protein structure, 2‐DE was used to determine the effect ofin vivolead exposure on GST isoform expression in rat kidney cytosols. Male Sprague‐Dawley rats were exposed to inorganic lead, and proteins were separated by conventional ISO‐DALT and NEPHGE‐DALT techniques and blotted for immunological identification. Lead exposure caused detectable inductions in both GSTP1 and GSTM1 and quantifiable charge modification in GSTP1. These preliminary data confirm the utility of 2‐D electrophoretic GST analysis as indicative of lead exposure and toxicity and support its use for further elaboration of lead's effects on renal pro

ISSN:0173-0835    

DOI:10.1002/elps.1150190821

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY

摘要:

AbstractIncorporation of the dispersion coefficient into the theory of the mobility‐shift assay for DNA‐protein complexes was highly successful largely due to increased mathematical rigor. A model simulating electrophoretic migration of DNA across the phase boundary between the initial zone of macromolecule and the gel lane predicts the peak asymmetry observed experimentally. It also predicts that, under the agency of the dispersion coefficient, the peak will become progressively more symmetrical during migration along the gel l

ISSN:0173-0835    

DOI:10.1002/elps.1150190822

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1998

数据来源: WILEY