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91.
F. W. van Glnkel W. B. van Muiswinkel ; B. Merchant E. F. Lizzio O. W. Dixon D. P. Anderson 《Journal of fish biology》1985,27(3):265-272
Anterior kidney and splenic cells were taken from rainbow trout and splenic cells from BALB/c mice immunized with a T-dependent (sheep red blood cells) or T-independent (DNP-Ficoll) antigen. The cells were incubated at different temperatures in Jerne plaque assays (direct or passive haemolytic plaque assays). The optimum numbers of in vitro plaque-forming cells (PFC) after incubation with homologous complement were directly correlated with normal body temperatures of the respective species. The optimum incubation temperature was 37°C for mouse cells and 10°C for fish cells. Incubation of mouse cells at lower temperatures of 30, 20, 10, 4 or 0°C appeared to yield a direct line reduction in numbers of PFC. Trout cells developed significantly fewer PFC at 4 and 20°C and none at 30°C or above; however, significant numbers still appeared at 0°C. More PFC per million white blood cells were obtained from the anterior kidney; however, related to temperatures, no differences in development of numbers of PFC could be seen between the spleen and anterior kidney cells of trout. When the incubation time was lengthened for both trout and mouse cells held at low temperatures, the numbers of PFC approached those of the cells incubated at the optimum temperatures for 10 h. 相似文献
92.
Homogenization of geographical variants at the nontranscribed spacer of rDNA in Drosophila mercatorum 总被引:2,自引:0,他引:2
rDNA nontranscribed spacer (NTS) lengths of Drosophila mercatorum have been
measured in individuals from several geographic regions. Individuals from
the different geographic subpopulations share some length fragments but are
in general distinct. The length differences, both within and between
individuals, arise from different copy numbers of a 250-bp repeating unit
that is localized to one part of the NTS. In addition to the length
differences caused by the 250-bp repeat, there is a Y chromosome
(male)-specific length variant elsewhere in the NTS that is approximately
70 bp shorter than the NTS fragment from the X chromosome. Sexual
dimorphism seems to be present in all Drosophila. Also, D. mercatorum has
fewer NTS length variants per individual than does D. melanogaster while
possessing comparable levels of restriction- site polymorphism. The
mechanisms that may cause this pattern of variation are selection, gene
conversion, and unequal recombination.
相似文献
93.
The influence of optimization target selection on the structure of arterial tree models generated by constrained constructive optimization 总被引:1,自引:1,他引:0
W Schreiner F Neumann M Neumann A End SM Roedler S Aharinejad 《The Journal of general physiology》1995,106(4):583-599
The computational method of constrained constructive optimization was used to generate complex arterial model trees by optimization with respect to a target function. Changing the target function also changes the tree structure obtained. For a parameterized family of target functions a series of trees was created, showing visually striking differences in structure that can also be quantified by appropriately chosen numerical indexes. Blood transport path length, pressure profile, and an index for relative segment orientation show clear dependencies on the optimization target, and the nature of changes can be explained on theoretical grounds. The main goal was to display, quantify, and explain the structural changes induced by different optimization target functions. 相似文献
94.
Naphthalene dioxygenase (NDO) fromPseudomonas sp strain NCIB 9816 is a multicomponent enzyme system which initiates naphthalene catabolism by catalyzing the addition of both atoms of molecular oxygen and two hydrogen atoms to the substrate to yield enantiomerically pure (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene. NDO has a relaxed substrate specificity and catalyzes the dioxygenation of many related 2- and 3-ring aromatic and hydroaromatic (benzocyclic) compounds to their respectivecis-diols. Biotransformations with a diol-accumulating mutant, recombinant strains and purified enzyme components have established that in addition tocis-dihydroxylation, NDO also catalyzes a variety of other oxidations which include monohydroxylation, desaturation (dehydrogenation),O-andN-dealkylation and sulfoxidation reactions. In several cases, the absolute stereochemistry of the oxidation products formed by NDO are opposite to those formed by toluene dioxygenase (TDO). The reactions catalyzed by NDO and other microbial dioxygenases can yield specific hydroxylated compounds which can serve as chiral synthons in the preparation of a variety of compounds of interest to pharmaceutical and specialty chemical industries. We present here recent work documenting the diverse array of oxidation reactions catalyzed by NDO. The trends observed in the oxidation of a series of benzocyclic aromatic compounds are compared to those observed with TDO and provide the basis for prediction of regio- and stereospecificity in the oxidation of related substrates. Based on the types of reactions catalyzed and the biochemical characteristics of NDO, a mechanism for oxygen activation by NDO is proposed. 相似文献
95.
Jesse C. Traller Shawn J. Cokus David A. Lopez Olga Gaidarenko Sarah R. Smith John P. McCrow Sean D. Gallaher Sheila Podell Michael Thompson Orna Cook Marco Morselli Artur Jaroszewicz Eric E. Allen Andrew E. Allen Sabeeha S. Merchant Matteo Pellegrini Mark Hildebrand 《Biotechnology for biofuels》2016,9(1):258
Background
Improvement in the performance of eukaryotic microalgae for biofuel and bioproduct production is largely dependent on characterization of metabolic mechanisms within the cell. The marine diatom Cyclotella cryptica, which was originally identified in the Aquatic Species Program, is a promising strain of microalgae for large-scale production of biofuel and bioproducts, such as omega-3 fatty acids.Results
We sequenced the nuclear genome and methylome of this oleaginous diatom to identify the genetic traits that enable substantial accumulation of triacylglycerol. The genome is comprised of highly methylated repetitive sequence, which does not significantly change under silicon starved lipid induction, and data further suggests the primary role of DNA methylation is to suppress DNA transposition. Annotation of pivotal glycolytic, lipid metabolism, and carbohydrate degradation processes reveal an expanded enzyme repertoire in C. cryptica that would allow for an increased metabolic capacity toward triacylglycerol production. Identification of previously unidentified genes, including those involved in carbon transport and chitin metabolism, provide potential targets for genetic manipulation of carbon flux to further increase its lipid phenotype. New genetic tools were developed, bringing this organism on a par with other microalgae in terms of genetic manipulation and characterization approaches.Conclusions
Functional annotation and detailed cross-species comparison of key carbon rich processes in C. cryptica highlights the importance of enzymatic subcellular compartmentation for regulation of carbon flux, which is often overlooked in photosynthetic microeukaryotes. The availability of the genome sequence, as well as advanced genetic manipulation tools enable further development of this organism for deployment in large-scale production systems.96.
Sequence of a cDNA for mouse thymidylate synthase reveals striking similarity with the prokaryotic enzyme 总被引:11,自引:0,他引:11
Perryman SM; Rossana C; Deng TL; Vanin EF; Johnson LF 《Molecular biology and evolution》1986,3(4):313-321
We report the nucleotide sequence of a cloned cDNA, pMTS-3, that contains a
1-kb insert corresponding to mouse thymidylate synthase (E.C. 2.1.1.45).
The open reading frame of 921 nucleotides from the first AUG to the
termination codon specifies a protein with a molecular mass of 34,962
daltons. The predicted amino acid sequence is 90% identical with that of
the human enzyme. The mouse sequence also has an extremely high degree of
similarity (as much as 55% identity) with prokaryotic thymidylate synthase
sequences, indicating that thymidylate synthase is among the most highly
conserved proteins studied to date. The similarity is especially pronounced
(as much as 80% identity) in the 44-amino-acid region encompassing the
binding site for deoxyuridylic acid. The cDNA sequence also suggests that
mouse thymidylate synthase mRNA lacks a 3' untranslated region, since the
termination codon, UAA, is followed immediately by a poly(A) segment.
相似文献
97.
Optimization of fermentation conditions for thermostable cellulase production byThielavia terrestris
Argyrios Margaritis Rosemina F. Merchant 《Journal of industrial microbiology & biotechnology》1986,1(3):149-156
Summary Of the eighteen different carbon sources, solka floc was optimal for the induction of cellulases by the thermophilic fungusThielavia terrestris. The temperature optimum for growth was between 44–52°C. The effect of initial and controlled pH on fungal growth and cellulase production was investigated and the results obtained showed that the maximum volumetric productivity (6.07 I.U./1 per h) of filter paper activity was achieved when the pH was controlled at 4.5–5.0. 相似文献
98.
Gary D. Stoner Donald J. Merchant 《In vitro cellular & developmental biology. Plant》1972,7(6):330-343
Summary The amino acid requirements of strain L-M mouse cells grown in a chemically defined medium (2×Eagle) containing only the 13
essential amino acids (EAA) were investigated. Medium and acid hydrolysate samples were analyzed for amino acid content by
the method of ion exchange chromatography. The extent of utilization of the EAA differed;e.g. after 120 hr of cell growth without medium change, glutamine was exhausted from the medium; methionine, leucine, isoleucine,
cystine, arginine, and valine were depleted 60 to 80%; other EAA were used to lesser extents. Although the EAA were used in
excess of their requirements for protein synthesis, a correlation could generally be made between utilization and protein
amino acid composition.
Glutamine appeared to be, a growth-limiting factor. Use of U-14C-labeled glutamine indicated that over one-half of the metabolized glutamine was converted to carbon dioxide, 17% to cell
material, and 15% was extracted from the amino acid pools.
Nonessential amino acids (NEAA), viz. alanine, aspartic acid, glutamic acid, glycine, proline, and serine, were released into
the medium during growth, and some were reutilized. Exogenous provision of these did not improve cell growth. In contrast
to the other NEAA, only serine showed net utilization when provided exogenously. When glutamic acid largely replaced the glutamine
in the medium, it exerted a sparing effect on the glutamine requirement for protein synthesis. Suggestions are given for the
improvement of Eagle medium for cell growth.
Supported by Research Grants CA 03720 and CA 11802 from the National Institutes of Health.
Predoctoral, fellow supported, by Grant F01-GM-42156-02 from the National Institutes of Health. 相似文献
99.
100.
VP4-specific intestinal antibody response to rotavirus in a murine model of heterotypic infection.
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We have adapted a murine model of heterotypic rotavirus infection for the purpose of evaluating the intestinal antibody response to an infection that mimics human vaccination. Neonatal mice were infected with the rhesus rotavirus (RRV). The enzyme-linked immunospot assay was used in order to avoid common artifacts in the quantitation of intestinal immune responses inherent in measurements of luminal or serum immunoglobulins and to obtain easily quantifiable data in a flexible and convenient format. Functionally active lymphocytes were harvested from the spleen, small intestinal lamina propria, Peyer's patches, and mesenteric lymph nodes and processed into single-cell suspensions. Antibody-secreting cells (ASC) were quantitated from 5 to 50 days after infection for total, RRV-specific, baculovirus-expressed VP4-specific, and single-shell RRV-specific ASC secreting either immunoglobulin G (IgG), IgM, or IgA. The response to VP4 constituted less than 1.5% of the total virus-specific response, which was located almost exclusively in the gut and was 90% IgA. Intestinal ASC were directed overwhelmingly toward proteins incorporated in the single-shell particle, predominantly VP2 and VP6. We conclude that the antibody response to VP4, thought to be the site of the important neutralization sites conserved among several rotavirus serotypes, is an extremely small portion of the overall antibody response in the intestinal tract. 相似文献