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51.
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
52.
A simple method for electrophoretic elution of nucleic acids from gel slices is described. The procedure utilizes a standard tube gel system and can be completed in as little as one hour. Nucleic acids are recovered in a small volume with almost 100% efficiency. The procedure is applicable equally to acrylamide and agarose gels, and small as well as large RNA and DNA molecules. The eluted nucleic acids are essentially undegraded and are suitable for a variety of structural and biological analyses. 相似文献
53.
Z. Krupa 《Photosynthesis research》1983,4(3):229-239
Thylakoid membranes obtained from bean chloroplasts treated with bean galactolipase or phospholipase A2 (from Crotalus terr. terr.) showed marked changes in their polypeptide patterns when separated on SDS-PAGE. The obtained results have been discussed with regard to the relationship between chloroplast lipids and polypeptides originating from chlorophyll-protein complexes of bean thylakoids. A coexistence between galactolipids and the peripheral antennae in PS I complex and LHCP3 as well as a conspicuous role of phospholipids in PSI and PSII centre chlorophyll-protein complexes has to be underlined.Abbreviations CP1
chlorophyll a-protein complex of PSI
- CPa
chlorophyll a-protein complex of PSII
- D10
digitonin subchloroplast particles enriched in PSII
- D144
digitonin subchloroplast particles enriched in PSI
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- LHCP1–3
light harvesting chlorophyll a/b protein complexes
- PAGE
polyacrylamide gel electrophoresis
- PSI
photosystem I
- PSII
photosystem II
- SDS
sodium dodecyl sulphate
- TCA
trichloroacetic acid
- Tricine
N-Tris-(hydroxymethyl)-methylglycine
- Tris
Tris-(hydroxymethyl)-aminomethan 相似文献
54.
55.
Stephen M. Beckstrom-Sternberg 《Biochemical Systematics and Ecology》1989,17(7-8):573-582
Two-dimensional polyacrylamide gel (2-D PAGE) electrophoresis was appraised as an experimental technique for assessing systematic relationships among higher plants and to determine at which level in the taxonomic hierarchy this technique is most generally applicable. 2-D PAGE was performed on denatured extracts of mature leaves from 25 species representing five families of the order Centrospermae (Caryophyllales, Chenopodiales) in the Angiospermae as well as Welwitschia mirabilis (Gymnospermae). Cluster analysis of a 256 spot binary-coded data set derived from the computer-encoded spot patterns of the 25 species successfully separated taxa from the individual to the familial levels of the taxonomic hierarchy in accordance with traditional taxonomic delineations of the taxa. 相似文献
56.
Characterization of differentiated syrian golden hamster pancreatic duct cells maintained in extended monolayer culture 总被引:4,自引:0,他引:4
S. Hubchak M. M. Mangino M. K. Reddy D. G. Scarpelli 《In vitro cellular & developmental biology. Plant》1990,26(9):889-897
Summary Epithelial cells isolated from fragments of hamster pancreas interlobular ducts were freed of fibroblast contamination by
plating them on air-dried collagen, maintaining them in serum-free Dulbecco's modified Eagle's (DME):F12 medium suppleneted
with growth factors, and selecting fibroblast-free aggregates of duct cells with cloning cylinders. Duct epithelial cells
plated on rat type I collagen gel and maintained in DME:F12 supplemented with Nu Serum IV, bovine pituitary extract, epidermal
growth factor, 3,3′, 5-triodothyronine, dexamethasone, and insulin, transferrin, selenium, and linoleic acid conjugated to
bovine serum albumin (ITS+), showed optimal growth as monolayers with a doubling time of about 20 h and were propagated for as long as 26 wk. Early
passage cells consisted of cuboidal cells with microvilli on their apical surface, complex basolateral membranes, numerous
elongated mitochondria, and both free and membrane-bound ribosomes. Cell grown as monolayers for 3 mo. were more flattened
and contained fewer apical microvilli, mitochondria, and profiles of rough surfaced endoplasmic reticulum; in addition, there
were numerous autophagic vacuoles. Functional characteristics of differentiated pancreatic duct cells which were maintained
during extended monolayer culture included intracellular levels of carbonic anhydrase and their capacity to generate cyclic
AMP (cAMP) after stimulation by 1×10−6
M secretin. From 5 to 7 wk in culture, levels of carbonic anhydrase remained stable but after 25 to 26 wk decreased by 1.9-fold.
At 5 to 7 wk of culture, cyclic AMP increased 8.7-fold over basal levels after secretin stimulation. Although pancreatic duct
cells cultured for 25 to 26 wk showed lower basal levels of cAMP, they were still capable of generating significant levels
of cAMP after exposure to serretin with a 7.0-fold increase, indicating that secretin receptors and the adenyl cyclase system
were both present and functional. These experiments document that pancreatic duct monolayer cultures can be maintained in
a differentiated state for up to 6 mo. and suggest that this culture system may be useful for in vitro physiologic and pathologic
studies.
This research was supported by grant CA34051 from the National Cancer Institute, Bethesda, MD. 相似文献
57.
Serum-free culture of enriched mouse anterior and ventral prostatic epithelial cells in collagen gel
Timothy Turner Howard A. Bern Peter Young Gerald R. Cunha 《In vitro cellular & developmental biology. Plant》1990,26(7):722-730
Summary Sustained growth of mouse ventral and anterior prostatic epithelial cells embedded within collagen gel matrix was achieved
in a serum-free medium composed of Dulbecco's modified Eagle's medium and Ham's F12 medium, 1∶1 (vol/vol), supplemented with
bovine serum albumin fraction V, epidermal growth factor, transferrin, cholera toxin, prolactin, 5α-dihydrotestosterone, cortisol,
putrescine, fibroblast growth factor, and a trace element mixture. Three-dimensional growth of prostatic epithelial cells
occurred inside the collagen gel matrix. This serum-free medium allowed cell growth greater than sevenfold over 10 d in culture.
Tissue recombination and cell culture techniques were integrated to demonstrate that cultured cells retained prostatic characteristics.
Following 10 d of culture, epithelial colonies from mouse ventral and anterior prostatic epithelial cell cultures were isolated
and combined with rat fetal urogenital sinus mesenchyme and grown for 4 wk under the renal capsule of intact athymic male
mice. These tissue recombinants showed distinctive prostatic histologic characteristic (alveoli and ducts lined with cuboidal
or columnar epithelium surrounded by stroma). When histologic sections of recombinants were stained with the Hoechst 33258,
epithelial cells of mouse origin were distinguishable from stromal cells of rat origin.
Aided by grants CA-05388 and CA-09041 from the National Institutes of Health, Bethesda, MD, and by M. A. R. C. fellowship
GM08730 to T. T. 相似文献
58.
Agar yield and quality of Gracilaria chilensis (Gigartinales,Rhodophyta) in tank culture using fish effluents 总被引:1,自引:0,他引:1
Tank cultivation of Gracilaria using fish effluents has permitted a production of 48 kg m–2 yr–1 and can reduce the dissolved nitrogen loads in the seawater. We report the yield, gel strength, gelling and melting point of agar from Gracilaria cultivated in tanks with seawater previously utilized in intensive, land-based salmon cultures and compared to a control using directly pumped seawater, over a study period of 22 months. The results show that the highest agar yield (20 to 22%) was obtained when Gracilaria was cultivated with pure seawater as compared to the fish effluents. The gel strength, gelling and melting point were higher in the agar obtained from algae cultured with fish effluents. During the spring, the gel strength, gelling and melting point increased in tanks with fish effluents and decreased in tanks with a supply of pure seawater. 相似文献
59.
A combined physical and genetic map of theCorynebacterium glutamicum ATCC 13032 chromosome was constructed using pulsed-field gel electrophoresis (PFGE) and hybridizations with cloned gene probes. Total genomic DNA was digested with the meganucleasesSwaI (5-ATTTAAAT-3),PacI (5-TTAATTAA-3), andPmeI (5-GTTTAAAC-3) yielding 26, 27, and 23 fragments, respectively. The chromosomal restriction fragments were then separated by PFGE. By summing up the lengths of the fragments generated with each of the three enzymes, a genome size of 3082 +/- 20 kb was determined. To identify adjacentSwaI fragments, a genomic cosmid library ofC. glutamicum was screened for chromosomal inserts containingSwaI sites. Southern blots of the PFGE gels were hybridized with these linking clones to connect theSwaI fragments in their natural order. By this method, about 90% of the genome could be ordered into three contigs. Two of the remaining gaps were closed by cross-hybridization of blottedSwaI digests using as probesPacI andPmeI fragments isolated from PFGE gels. The last gap in the chromosomal map was closed by hybridization experiments using partialSwaI digestions, thereby proving the circularity of the chromosome. By hybridization of gene probes toSwaI fragments separated by PFGE about 30 genes, including rRNA operons, IS element and transposon insertions were localized on the physical map. 相似文献
60.
Hisao Kurazono Shinji Yamasaki Orn-anong Ratchtrachenchai G. Balakrish Nair Yoshifumi Takeda 《Microbiology and immunology》1996,40(4):303-305
Vibrio cholerae O139 isolated from different countries, as well as from different locations within a country, were examined using macrorestriction DNA analysis to determine the clonality of the O139 strains. NotI digests of genomic DNA of representative strains from Nepal, India, Bangladesh, China, Thailand, and Malaysia revealed very similar but not identical patterns. Examinations of the banding patterns generated by pulsed-field gel electrophoresis of strains isolated within countries revealed complete homogeneity. These results further reiterate the spread of an identical clone of V. cholerae O139 although it appears that genetic polymorphism among the O139 strains is becoming apparent. 相似文献