摘要:

AbstractAs the first step in a cooperative effort to standardize the identification of the polypeptides ofTreponema pallidumsubsp.pallidum, the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) results obtained in 16 laboratories were compared. Although it was possible to correlate the positions of 16 of the major polypeptide bands, the cross‐identification of many of the polypeptides was ambiguous, particularly in the low molecular weight range. Two‐dimensional electrophoresis provided an improved means of separating and characterizingT. pallidumpolypeptides as isolated molecular species. An approach to the unambiguous identification of treponemal polypeptides was outlined which will utilize two‐dimensional electrophoresis in combination with specific properties attributable to individual proteins, including reactivity with monoclonal antibodies or monospecific antisera, biochemical and structural properties, and sequence information. To demonstrate the feasibility of this approach, two‐dimensional electrophoresis in conjunction with immunoperoxidase staining was used to specifically identify three clonedT. palli

ISSN:0173-0835    

DOI:10.1002/elps.1150080202

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

年代:1987

数据来源: WILEY

摘要:

AbstractA new modification of silver staining is presented which utilizes two chemical properties of thiosulfate: image enhancement by pretreatment of fixed gels, and formation of soluble silver complexes which prevents unspecific background staining during image development. This procedure provides high sensitivity for proteins, RNA and DNA in the nanogram range on a colorless, transparent background. The performance of this method is documented by staining one‐and two‐dimensional patterns of plant leaf proteins. Moreover, we achieved, for the first time, the detection of the non‐structural, tobacco mosaic virus‐specific 126 kDa protein directly in the one‐dimensional protein pattern of infected protoplasts by a staining

ISSN:0173-0835    

DOI:10.1002/elps.1150080203

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

年代:1987

数据来源: WILEY

摘要:

AbstractOne of the most critical steps in two‐dimensional gel analysis is the comparison of spot lists. In this paper, we describe our approach to the matching problem. We propose a method based on syntactic pattern recognition techniques, which differs from previously used methods and offers a major improvement to cope with discontinuous geometric distortions. The method is based on non‐metric considerations and describes the pattern observed on a given gel in a non‐geometric way. This approach leads to the definition of finite automatons which can be used to automatically find the presence of a given pattern on a gel. The program uses a technique of clause generation related to artificial intelligence techniques. A set of rules is applied to the definition of homologousspots, and probability coefficients are computed to compare and to chose between different pairing hypotheses. Once matching has been achieved between a set of gels, the construction of a master gel which summarizes the information obtained is allowed. Two types of masters can be used. The first one summarizes the information between spots lists belonging to the same experiment and includes intensity considerations, while the second one summarizes the information resulting from different gels and does not include spot intensity and volume. Masters are written and known by the system as a spot list. This ability allows them to be handled just like spot lists and, using them, to perform all the actions available to the system. This approach is compared with a more classical one based on geometric corrections. The use of probability coefficients is also discussed and extended to their application to multiple matching and further utilization of ma

ISSN:0173-0835    

DOI:10.1002/elps.1150080204

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

年代:1987

数据来源: WILEY

摘要:

AbstractIn the androgenized rat, uterine histological abnormalities associated with unopposed oestrogenic stimulation can be partially reversed following progestin therapy. The effects of progestins and tamoxifen on the uterine protein composition of androgenized animals have therefore been investigated in an attempt to identify hormonally sensitive proteins that reflect changes in tissue morphology. Subcellular fractions prepared from uteri of androgenized animals were analysed by one‐ and two‐dimensional gel electrophoresis. In all fractions, protein changes were observed and the identity of some of these proteins has been tentatively assigned following peptide mapping. Evidence is presented of an apparent correlation between potency to antagonize the effects of unopposed oestrogenic stimulation and an increase in the proportion of cytosolic glucose‐6‐phosphate dehydrogenase and the B form of creatine

ISSN:0173-0835    

DOI:10.1002/elps.1150080205

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

年代:1987

数据来源: WILEY

摘要:

AbstractGenetic polymorphism of human lymphocyte proteins was studied by two‐dimensional electrophoresis in the members of 12 families and in additional 20 unrelated individuals from Southern Germany. Cell lysates of phytohemagglutinin‐stimulated, [3H‐]leucine labeled peripheral lymphocytes were subjected to two‐dimensional electrophoresis. Among the approximate 300 labeled proteins on two‐dimensional fluorograms we found six polymorphic proteins. Five proteins, named p23 (Mr23 000), p30 (Mr30 000), p40 (Mr40 000), p60 (Mr60 000) and p66 (Mr66 000), existed as two charge variants, an acidic (a) and a basic (b) variant. Three different electrophoretic phenotypes, two homozygous (a and b) and one heterozygous (ab), could be observed for the proteins p23, p40 and p66. The proteins p30 and p60 occurred in our population sample only in two phenotypes, a common homozygous (a) and a rare heterozygous (ab) phenotype. Preliminary results are presented for the sixth polymorphic protein p75 (Mr75 000), which occurred in at least three or even more different electrophoretic variants, resulting in six or more phenotypes. The genetic basis of the protein variations was confirmed by the quantitative gene‐dosage dependence, the identity of the protein patterns in three monozygotic twin pairs and the transmission of the variants as Mendelian traits within the 12 families examined. Gene frequencies of the rare alleles of the five proteins which occurred in two charge variants ranged between 0.012 and 0.477 in a sample of 44 unrelated individuals from Southern Germany. Thus, the term genetic polymorphism i

ISSN:0173-0835    

DOI:10.1002/elps.1150080206

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

年代:1987

数据来源: WILEY

摘要:

AbstractSilver staining artifacts, unrelated to protein, such as horizontal lines and vertical micro or point streaking on two‐dimensional gels with carrier ampholytes or immobilized pH gradients in the first dimension are efficiently eliminated by adding iodoacetamide to the equilibration buffer of the first‐dimensional

ISSN:0173-0835    

DOI:10.1002/elps.1150080207

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150080208

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150080201

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

年代:1987

数据来源: WILEY