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51.
Organization of the ribosomal ribonucleic acid genes in various wild-type strains and wild-collected strains of Neurospora 总被引:6,自引:0,他引:6
Peter J. Russell Sheryl Wagner Karin D. Rodland Rhonda L. Feinbaum Jennifer P. Russell Marion S. Bret-Harte Stephen J. Free Robert L. Metzenberg 《Molecular & general genetics : MGG》1984,196(2):275-282
Summary The organization of the ribosomal DNA (rDNA) repcat unit in the standard wild-type strain of Neurospora crassa, 74-OR23-1A, and in 30 other wild-type strains and wild-collected strains of N. crassa, N. tetrasperma, N. sitophila, N. intermedia, and N. discreta isolated from nature, was investigated by restriction enzyme digestion of genomic DNA, and probing of the Southern-blotted DNA fragments with specific cloned pieces of the rDNA unit from 74-OR23-1A. The size of the rDNA unit in 74-OR23-1A was shown to be 9.20 kilobase pairs (kb) from blotting data, and the average for all strains was 9.11+0.21 kb; standard error=0.038; coefficient of variation (C.V.)=2.34%. These data indicate that the rDNA repeat unit size has been highly conserved among the Neurospora strains investigated. However, while all strains have a conserved HindIII site near the 5 end of the 25 S rDNA coding sequence, a polymorphism in the number and/or position of HindIII sites in the nontranscribed spacer region was found between strains. The 74-OR23-1A strain has two HindIII sites in the spacer, while others have from 0 to at least 3. This restriction site polymorphism is strain-specific and not species-specific. It was confirmed for some strains by restriction analysis of clones containing most of the rDNA repeat unit. The current restriction map of the 74-OR23-1A rDNA repeat unit is presented. 相似文献
52.
Jennifer Allsop Richard W.E. Watts 《Differentiation; research in biological diversity》1986,32(2):144-147
The overall activity of the purine de novo synthesis pathway and the activities of purine phosphoribosyltransferase in the rat testis were measured at different ages and were correlated with histological observations. Similar studies of the concentration of circulating gonadotrophins and testosterone were performed. The purine phosphoribosyltransferase activities were between two and three orders of magnitude greater than purine de novo synthesis. The peak activity of the purine de novo synthesis pathway coincided with the first appearance of meiosis in the spermatocytes immediately before the luteinising hormone (LH) level rose to its peak. The highest activity of the hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8) - catalysed purine salvage pathway coincided with the first appearance of mature spermatozoa in the tubules just after the occurrence of peak levels of follicle-stimulating hormone (FSH). These findings are linked to the development of testicular atrophy in cases of severe HPRT deficiency in man. 相似文献
53.
Bovine trypsinogen and chymotrypsinogen were successfully refolded as the mixed disulfide of glutathione using cysteine as the disulfide interchange catalyst. The native structures were regenerated with yields of 40%-50% at pH 8.6 and 4 degrees C, and the half-time for the refolding was approximately 60-75 min. We then refolded threonine-neochymotrypsinogen, which is a two-chain structure held together by disulfide bonds and produced on cleavage of Tyr 146-Thr 147 in native chymotrypsinogen [Duda CT, Light A, J Biol Chem 257 9866-9871, 1982]. Neochymotrypsinogen was denatured and fully reduced, and the thiols were converted to the mixed disulfide of glutathione. The two polypeptide fragments, representing the amino- and carboxyl-terminal domains, were separated on Sephadex G-75. Mixtures of the polypeptide fragments varying in the ratio of their concentration from 1:5 to 5:1 were refolded with yields of 21-28%. The lack of dependence on the concentration of either fragment and the relatively high yields suggest independent folding of the amino- and carboxyl-terminal domains. When the globular structures of the domains formed, they then interacted with one another and produced the native intermolecular disulfide bridge and the proper geometry of the active site. 相似文献
54.
Mavis Hendson Lizzie Askjaer Jennifer A. Thomson Marc van Montagu 《Applied microbiology》1983,45(5):1526-1532
Eighteen strains of Agrobacterium tumefaciens isolated from crown galls were tested for agrocin production. Of six agrocin-producing strains, one (D286) produced a broad-host-range agrocin active against strains carrying nopaline, octopine, and agropine type Ti plasmids. Sensitivity to agrocin D286 was found to map in the 11- to 18-megadalton region of the nopaline Ti plasmid pTiC58. The agrocin was partially purified, and its physical characteristics were consistent with its being a nucleotide, as is agrocin 84. Agrocin D286 was shown to inhibit DNA, RNA, and protein syntheses. Strain D286 spontaneously lost its pathogenicity, and its potential for use in the biological control of crown gall is discussed. 相似文献
55.
1. Mycophenolic acid, an antibiotic of some antiquity that more recently has been found to have marked activity against a range of tumours in mice and rats, strongly inhibits DNA synthesis in the L strain of fibroblasts in vitro. 2. The extent of the inhibition of DNA synthesis is markedly increased by preincubation of the cells with mycophenolic acid before the addition of [(14)C]thymidine. 3. The inhibition of DNA synthesis by mycophenolic acid in L cells in vitro is reversed by guanine in a non-competitive manner, but not by hypoxanthine, xanthine or adenine. 4. The reversal of inhibition by guanine can be suppressed by hypoxanthine, 6-mercaptopurine and adenine. 5. Mycophenolic acid does not inhibit the incorporation of [(14)C]thymidine into DNA in suspensions of Landschütz and Yoshida ascites cells in vitro. 6. Mycophenolic acid inhibits the conversion of [(14)C]hypoxanthine into cold-acid-soluble and -insoluble guanine nucleotides in Landschütz and Yoshida ascites cells and also in L cells in vitro. There is some increase in the radioactivity of the adenine fraction in the presence of the antibiotic. 7. Mycophenolic acid inhibits the conversion of [(14)C]hypoxanthine into xanthine and guanine fractions in a cell-free system from Landschütz cells capable of converting hypoxanthine into IMP, XMP and GMP. 8. Preparations of IMP dehydrogenase from Landschütz ascites cells, calf thymus and LS cells are strongly inhibited by mycophenolic acid. The inhibition showed mixed type kinetics with K(i) values of between 3.03x10(-8) and 4.5x10(-8)m. 9. Evidence was also obtained for a partial, possibly indirect, inhibition by mycophenolic acid of an early stage of biosynthesis of purine nucleotides as indicated by a decrease in the accumulation of formylglycine amide ribonucleotide induced by the antibiotic azaserine in suspensions of Landschütz and Yoshida ascites cells and L cells in vitro. 相似文献
56.
Toxin Production in Clostridium botulinum as Demonstrated by Electron Microscopy 总被引:1,自引:0,他引:1 下载免费PDF全文
Spheroplasts of Clostridium botulinum 62A were prepared with the use of lysozyme. These spheroplasts were then exposed to ferritin-labeled type A antitoxin. Ultrathin sections of these specimens revealed the ferritin-labeled antibody symmetrically arranged around the outer spore coats but not within the spore cortex. The ferritin-labeled antibody was also observed in the bacterial cytoplasm. Here it was arranged in aggregates and strands, although it was not associated with any identifiable cell structure. Controls included sections of C. botulinum spheroplasts treated with a 1.5% solution of ferritin as well as spheroplasts of C. roseum and Bacillus subtilis treated with conjugated type A antitoxin or a 1.5% solution of ferritin. No intracellular or extracellular ferritin was demonstrable in these specimens. 相似文献
57.
Summary The ultrastructure of the haematopoietic cells in the polychaetes Neoamphitrite figulus Dalyell, Lanice conchilega (Pallas), Arenicola marina (L.), Myxicola infundibulum Renier, Megalomma vesiculusom (Montagu), Sabella penicillus L., are compared: all show similarities in having well developed Golgi, granular endoplasmic reticulum and haemoglobin or chlorocruorin in vesicles, and numerous mitochondria. The porphyrin byproducts of synthesis are combined with iron as haematins within electrondense granules built up from multi-lamellar organelles. The structure of the basal lamina which alone separates the cells from the lumen of the vessels is described and evidence is presented for the method of release of the haem into the plasma by reverse pinocytosis. The cycle of synthesis within the cell is discussed and the process of haem synthesis in annelids is reviewed. The structure of the haemoglobin-containing coelomocytes of Neoamphitrite figulus is briefly described.This work was made possible by an award from the Science Research Council. We would also like to thank the Director and Staff of the Plymouth Laboratory, where some of the work was done, for their hospitality and facilities. 相似文献
58.
59.
Hillary A. Hahm Margot M. Ip Kathleen Darcy Jennifer D. Black Wendy K. Shea Suzanne Forczek Masami Yoshimura Takami Oka 《In vitro cellular & developmental biology. Plant》1990,26(8):803-814
Summary A serum-free primary culture system is described which allows normal rat mammary epithelial cells (RMECs) embedded within
a reconstituted basement membrane to undergo extensive growth and functional differentiation as detected by synthesis and
secretion of the milk products casein and lipid. RMECs isolated from mammary glands of immature virgin rats were seeded within
an extracellular matrix preparation derived from the Engelbreth-Holm-Swarm sarcoma and cultured in a serum-free medium consisting
of Dulbecco's modified Eagle's medium-F12 containing insulin, prolactin, progesterone, hydrocortisone, epidermal growth factor,
bovine serum albumin, transferrin, and ascorbic acid. Casein synthesis and secretion were documented at the electron microscopic
level as well as by an enzyme-linked immunosorbent assay (ELISA) assay using a polyclonal antibody against total rat caseins.
Numerous secretory vesicles with casein micelles were noted near the apical surface of the RMECs, and secreted casein was
observed in the lumen. These ultrastructural data were confirmed by the ELISA assay which showed that microgram amounts of
casein per well were synthesized by the RMECs and that the amount of casein increased with time in culture. Using immunoblot
analysis it was demonstrated that the full complement of casein proteins was synthesized. In addition to casein protein, β-casein
mRNA levels were shown to increase with time. Synthesized lipid was detected at both the light and electron microscopic levels.
Phase contrast photomicrographs demonstrated extensive intracellular lipid accumulation within the ductal and lobuloalveolarlike
colonies, and at the electron micrograph level, lipid droplets were predominantly localized near the apical surface of the
RMECs. The lipid nature of these droplets was verified by oil red O staining. Results from this study demonstrate that RMECs
from immature virgin rats proliferate extensively and rapidly develop the capacity to synthesize and secrete casein and lipid
when grown within a reconstituted basement membrane under defined serum-free conditions. This unique system should thus serve
as an excellent model in which the regulation of mammary development and gene expression can be investigated.
This work was supported by grants CA 33240 and CA 35641 and by core grant CA 24538 from the National Institutes of Health,
Bethesda, MD. 相似文献
60.
Michael A. Bowers Stephen F. Matter James L. Dooley Jr. Jennifer L. Dauten John A. Simkins 《Oecologia》1996,108(1):182-191
Habitat fragmentation involves a reduction in the effective area available to a population and the imposition of hard patch edges. Studies seeking to measure effects of habitat fragmentation have compared populations in fragments of different size to estimate and area effect but few have examined the effect of converting open populations to closed ones (an effect of edges). To do so requires a shift in spatial scope-from comparison of individual fragments to that of fragmented versus unfragmented landscapes. Here we note that large-scale, controlled studies of habitat fragmentation have rarely been performed and are needed. In making our case we develop a simple computer simulation model based on how individual animals with home ranges are affected by the imposition of habitat edges, and use it to predict population-level responses to habitat fragmentation. We then compare predictions of the model with results from a field experiment on Peromyscus and Microtus. Our model treats the case where home ranges/territories fall entirely within or partially overlap with that of sample areas in continuous landscapes, but are restricted to areas within habitat fragments in impacted landscapes. Results of the simulations demonstrate that the imposition of hard edges can produce different population abundances for similar-sized areas in continuous and fragmented landscapes. This edge effect is disproportionately greater in small than large fragments and for species with larger than smaller home ranges. These predictions were generally supported by our field experiment. We argue that large-scale studies of habitat fragmentation are sorely needed, and that control-experiment contrasts of fragmented and unfragmented microlandscapes provide a logical starting point. 相似文献