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21.
An electrophoretic procedure for separating the molecular forms of catechol-O-methyltransferase in cellulose acetate gel is described; the zones of enzyme activity were revealed by autoradiography. The electrophoretic patterns of the enzyme in several tissues and cell lines derived from four different species are presented.This investigation was supported by Grant 500.6/Ric. 70/1981 from the Italian Ministry of Health and by the Italian Ministry of Education. We are grateful to Dr. M. Castagnola for useful advice and help with thin-layer chromatography. 相似文献
22.
Electrophoretic analysis of polyphenoloxidase isoenzymes from a variety of angiosperms and from mushroom revealed that the enzymes remain active in the presence of 0.1 % sodium dodecylsulfate. Electrophoresis in the presence of sodium dodecylsulfate allows the detection of latent enzyme forms of polyphenoloxidase, and can also convert slower migrating enzyme forms to faster migrating forms. Electrophoresis in the absence of sodium dodecylsulfate followed by incubation in the presence of sodium dodecylsulfate can also be used to detect latent forms of polyphenoloxidase. Together, these approaches provide a method for screening latent enzymes and give some insight into the mechanism of activation by sodium dodecylsulfate. 相似文献
23.
Y Yamada 《Journal of biochemical and biophysical methods》1983,8(3):175-181
An apparatus has been developed to reduce cathodic drift and migration into the anode chamber in vertical gel rod isoelectric focusing (IEF). In contrast to commercially available apparatuses, this apparatus can easily handle many more gels at one time, and the length, diameter and shape of its gel can be arbitrarily changed. In addition, high concentrations of detergent can be used to dissolve the protein samples, and removal of the gel cylinders from the glass tubes is easy. 相似文献
24.
Wilfried Bautsch 《Molecular biotechnology》1994,2(1):29-44
Macrorestriction mapping is often the first step toward a thorough physical and genetic characterization of a bacterial genome.
The problem of deducing the order of partially or completely digested macrorestriction fragments to yield a physical genome
map may readily be solved by applying twodimensional pulsed-field gel electrophoresis (2D-PFGE) techniques. These powerful
methods are quick and technically easy to perform; specifically, they are independent of DNA probes and should therefore be
applicable to any bacterial species irrespective of its prior genetic characterization. In this article, detailed step-by-step
protocols are given to set up, run, and evaluate 2D pulsed-field gels. Two basic methods are described: partial/complete 2D
gels of one restriction enzyme and complete/complete 2D gels of two different restriction enzymes. Other topics include preparation
of bacterial genomic DNA, screening for suitable rare-cutting restriction enzymes and determination of optimal running conditions.
Accompanied by many notes, these protocols are meant to offer the novice a sound and rapid access to these important methods. 相似文献
25.
DAVID GOULSON 《Biological journal of the Linnean Society. Linnean Society of London》1993,49(2):127-139
Most temperate butterflies exhibit a tightly synchronized unimodal adult emergence to facilitate mate location. Exceptions are presumably subject to unusual selection pressure. This study examines the pattern of emergence in Maniola jurtina , which was found to exhibit both unimodal and bimodal emergence patterns at different sites in south-east England. The bimodal pattern was found on chalk grassland; elsewhere the emergence was unimodal. Adults from each emergence peak rarely meet, suggesting that there may be some degree of reproductive isolation. Morphological measurements and electrophoretic analysis of allozyme frequencies are carried out to quantify differentiation between emergence peaks. Captive stock was reared to examine differences in the immature stages. Butterflies from each emergence differ significantly in most morphological variables measured, those from the second peak tending to be smaller. The immature stages differ in morphology and longevity of the egg stage. Allozyme frequencies did not differ between peaks, suggesting that they are not reproductively isolated. Explanations for the maintenance of differences between emergence peaks despite gene flow are discussed. I propose that division of offspring between two emergence times may have evolved to avoid the risk inherent in placing all offspring in one peak which may be rendered inviable by temporal fluctuations in habitat quality. 相似文献
26.
27.
The demonstration of prophenoloxidase after electrophoresis is based on its activation by sodium dodecyl sulfate (SDS) or sodium oleate and staining the activated phenoloxidase with dopamine and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH). A rapid method is presented for demonstrating the presence of activated phenoloxidase using polyacrylamide gel electrophoresis followed by staining in the presence of SDS or sodium oleate. 相似文献
28.
Yukie Takahashi Masako Sohnaka Minoru Nakao Kenji Miyamoto Masahito Fukunaga 《Microbiology and immunology》1993,37(9):721-727
Thirty-two Borrelia isolates were obtained from the adult stage of ixodid ticks, Ixodes ovatus, collected in various localities in Japan. Borrelial isolates were cultivated and analyzed by polyacrylamide gel electrophoresis, with monoclonal antibodies, by pulsed field gel electrophoresis, and by genomic Southern hybridization. All borrelial isolates showed similar protein profiles and monoclonal antibody reactivities, while plasmid profiles were rather diverse. Genomic hybridization using rRNA gene probes demonstrated the genetic similarities of those isolates. We found no significant differences among the borrelial isolates tested, and the restriction fragment length polymorphism patterns of I. ovatus isolates were quite distinct from those of borrelial strains associated with Lyme disease. Therefore, the isolates of Borrelia obtained from I. ovatus were thought to fall into different genospecies. 相似文献
29.
Maria J. P. van Dongen Sybren S. Wijmenga Ramón Eritja Fernando Azorín Cornelis W. Hilbers 《Journal of biomolecular NMR》1996,8(2):207-212
Summary A new application of the HMBC experiment is presented that provides a useful means to discriminate between H2 and H8 proton resonances, to assign the base proton resonances to the various residue types and, most importantly, to correlate the H2 and H8 protons for adenine or inosine residues in natural abundance 13C fragments. The utility of this experiment is demonstrated for an unlabeled DNA 20-mer. Thanks to the obtained results, preliminary conclusions could be drawn regarding the molecular conformations of the non-canonical G/I-A base pairs in the hairpin formed by this fragment. 相似文献
30.
A novel approach to the analysis of the initiation of embryo development in gramineae 总被引:1,自引:1,他引:0
F. Matzk H.-M. Meyer H. Bäumlein H.-J. Balzer I. Schubert 《Sexual plant reproduction》1995,8(5):266-272
An in vivo model system to study the initiation of embryo development is presented. From the so-called Salmon system of wheat (alloplasmic lines with a 1BL-1RS chromosome translocation), three completely isogenic and homozygous lines were produced by selection for uniformity in about 20 selfing/backcross generations as well as between sublines of doubled haploids. The line (aestivum)-Salmon is male fertile and sexual. The lines (caudata)-Salmon and (kotschyi)-Salmon are male sterile and have a parthenogenetic capacity of about 90%. The expression of nuclear-cytoplasmic male sterility is different for the two parthenogenetic lines. The initiation of autonomous embryo development at defined developmental stages of the ovaries and the maximum degree of parthenogenesis are identical in both parthenogenetic lines as proved by the auxin test and progeny analyses. The protein patterns from ovary extracts of the three isogenic lines were identical for more than 200 spots of 2-D polyacrylamide gels, confirming their homogeneity. However, one protein (P 115.1) was found 3 days before and during anthesis only in ovaries of the parthenogenetic lines. It seems to be involved in the initiation of parthenogenesis. 相似文献