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41.
Tallury SP Hilu KW Milla SR Friend SA Alsaghir M Stalker HT Quandt D 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,111(7):1229-1237
Section Arachis is the largest of nine sections in the genus Arachis and includes domesticated peanut, A. hypogaea L. Most species are diploids (x=10) with two tetraploids and a few aneuploids. Three genome types have been recognized in this section (A, B and D), but
the genomes are not well characterized and relationships of several newly described species are uncertain. To clarify genomic
relationships in section Arachis, cytogenetic information and molecular data from amplified fragment length polymorphism (AFLP) and the trnT-F plastid region were used to provide an additional insight into genome composition and species relationships. Cytogenetic
information supports earlier observations on genome types of A. cruziana, A. herzogii, A. kempff-mercadoi and A. kuhlmannii but was inconclusive about the genome composition of A. benensis, A. hoehnei, A. ipaensis, A. palustris, A. praecox and A. williamsii. An AFLP dendrogram resolved species into four major clusters and showed A. hypogaea grouping closely with A. ipaensis and A. williamsii. Sequence data of the trnT-F region provided genome-specific information and showed for the first time that the B and D genomes are more closely related
to each other than to the A genome. Integration of information from cytogenetics and biparentally and maternally inherited
genomic regions show promise in understanding genome types and relationships in Arachis. 相似文献
42.
The simple repeat poly(dT-dG).poly(dC-dA) common to eukaryotes is absent from eubacteria and archaebacteria and rare in protozoans 总被引:5,自引:0,他引:5
Genomic DNA from a wide variety of prokaryotic and eukaryotic organisms has
been assayed for the simple repeat sequence poly(dT-dG).poly(dC-dA) by
Southern blotting and DNA slot blot hybridizations. Consistent with
findings of others, we have found the simple alternating sequence to be
present in multiple copies in all organisms in the animal kingdom (e.g.,
mammals, reptiles, amphibians, fish, crustaceans, insects, jellyfish,
nematodes). The TG element was also found in lower eukaryotes
(Saccharomyces cerevisiae, Neurospora crassa, and Dictyostelium discoideum)
and at a much lower frequency in protozoans (Oxytricha fallux and
Tetrahymena thermophila). The sequence was also repeated in high copy
number in a higher plant (Zea mays) as well as at very high levels in a
unicellular green alga (Chlamydomonas reinhardi). Although the copy number
of the repeat per haploid genome was generally proportional to genome size,
there was a greater-than-1,000-fold variation in the number of
(TG)25/100-kb genomic DNA. By contrast, no eu-or archaebacterium--including
Myxococcus xanthus, whose life cycle is very similar to that of the slime
mold Dictyostelium discoideum, and Halobacter volcanii, whose genome
contains other repeated sequences-- was found whose genomic DNA contained
this sequence in detectable amounts. A computer search also failed to find
the TG element in human mitochondrial DNA.
相似文献
43.
S R Padgette Q K Huynh S Aykent R D Sammons J A Sikorski G M Kishore 《The Journal of biological chemistry》1988,263(4):1798-1802
Reaction of 5-enolpyruvylshikimate-3-phosphate synthase of Escherichia coli with the thiol reagent 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) leads to a modification of only 2 of the 6 cysteines of the enzyme, with a significant loss of its enzymatic activity. Under denaturing conditions, however, all 6 cysteines of 5-enolpyruvylshikimate-3-phosphate synthase react with DTNB, indicating the absence of disulfide bridges in the native protein. In the presence of shikimate 3-phosphate and glyphosate, only 1 of the 2 cysteines reacts with the reagent, with no loss of activity, suggesting that only 1 of these cysteines is at or near the active site of the enzyme. Cyanolysis of the DTNB-inactivated enzyme with KCN leads to elimination of 5-thio-2-nitrobenzoate, with formation of the thiocyano-enzyme. The thiocyano-enzyme is fully active; it exhibits a small increase in its I50 for glyphosate (6-fold) and apparent Km for phosphoenolpyruvate (4-fold) compared to the unmodified enzyme. Its apparent Km for shikimate 3-phosphate is, however, unaltered. These results clearly establish the nonessentiality of the active site-reactive cysteine of E. coli 5-enolpyruvylshikimate-3-phosphate synthase for either catalysis or substrate binding. Perturbations in the kinetic constants for phosphoenolpyruvate and glyphosate suggest that the cysteine thiol is proximal to the binding site for these ligands. By N-[14C]ethylmaleimide labeling, tryptic mapping, and N-terminal sequencing, the 2 reactive cysteines have been identified as Cys408 and Cys288. The cysteine residue protected by glyphosate and shikimate 3-phosphate from its reaction with DTNB was found to be Cys408. 相似文献
44.
Steven Slater Kathryn L. Houmiel Minhtien Tran Timothy A. Mitsky Nancy B. Taylor Stephen R. Padgette Kenneth J. Gruys 《Journal of bacteriology》1998,180(8):1979-1987
Polyhydroxyalkanoates (PHAs) are a class of carbon and energy storage polymers produced by numerous bacteria in response to environmental limitation. The type of polymer produced depends on the carbon sources available, the flexibility of the organism’s intermediary metabolism, and the substrate specificity of the PHA biosynthetic enzymes. Ralstonia eutropha produces both the homopolymer poly-β-hydroxybutyrate (PHB) and, when provided with the appropriate substrate, the copolymer poly(β-hydroxybutyrate-co-β-hydroxyvalerate) (PHBV). A required step in production of the hydroxyvalerate moiety of PHBV is the condensation of acetyl coenzyme A (acetyl-CoA) and propionyl-CoA to form β-ketovaleryl-CoA. This activity has generally been attributed to the β-ketothiolase encoded by R. eutropha phbA. However, we have determined that PhbA does not significantly contribute to catalyzing this condensation reaction. Here we report the cloning and genetic analysis of bktB, which encodes a β-ketothiolase from R. eutropha that is capable of forming β-ketovaleryl-CoA. Genetic analyses determined that BktB is the primary condensation enzyme leading to production of β-hydroxyvalerate derived from propionyl-CoA. We also report an additional β-ketothiolase, designated BktC, that probably serves as a secondary route toward β-hydroxyvalerate production.Polyhydroxyalkanoates (PHAs) are a class of naturally occurring polymers which serve as a carbon and energy reserve in numerous bacterial species. Ralstonia eutropha (formerly designated Alcaligenes eutrophus [41]) produces the homopolymer poly(β-hydroxybutyrate) (PHB) and, when provided with propionate in the feedstock, the copolymer poly(β-hydroxybutyrate-co-β-hydroxyvalerate) (PHBV). R. eutropha is used commercially to produce PHBV, which is a biodegradable thermoplastic.The PHB biosynthetic pathway requires three enzymatic activities: a β-ketothiolase (PhbA), an NADPH-dependent acetoacetyl coenzyme A (acetoacetyl-CoA) reductase (PhbB) and a PHB synthase (PhbC). The first step in production of the homopolymer PHB is catalyzed by β-ketothiolase which condenses two acetyl-CoA molecules to form acetoacetyl-CoA. Formation of the copolymer PHBV requires the additional condensation of acetyl-CoA with propionyl-CoA to form β-ketovaleryl-CoA (Fig. (Fig.1).1). Subsequently, the acetoacetyl-CoA and β-ketovaleryl-CoA are converted into a polymer by the activities of the reductase and synthase. The genes encoding these proteins in R. eutropha reside in an operon which has been well characterized (10, 21, 22, 31, 37). Open in a separate windowFIG. 1Pathway for production of PHBV from acetyl-CoA and propionyl-CoA. β-Ketothiolase performs the condensation reactions to generate either acetoacetyl-CoA or β-ketovaleryl-CoA. These are reduced by acetoacetyl-CoA reductase (PhbB) and polymerized by PHB synthase (PhbC).The substrate specificities of these three enzymes are reportedly adequate for production of PHBV copolymer (7–9), but propionate-fed Escherichia coli harboring the R. eutropha phb operon produces essentially PHB homopolymer (35). Moreover, PHBV copolymer can be produced in E. coli after induction of the fatty acid β-oxidation complex, which contains a β-ketothiolase with broad substrate specificity (26, 27, 35). These data suggest that the R. eutropha PHB pathway is capable of producing copolymer, but only in the context of a second β-ketothiolase with broad substrate specificity.R. eutropha is known to produce at least two β-ketothiolases (7), and at least two distinct plasmid clones which express β-ketothiolase have been isolated from R. eutropha (37). In this work, we analyzed the substrate specificity of the PhbA β-ketothiolase and demonstrated that this enzyme catalyzes thiolysis of β-ketovaleryl-CoA very poorly. We determined that R. eutropha expresses at least two β-ketothiolases in addition to PhbA and that these additional enzymes, which we designate BktB and BktC, efficiently utilize β-ketovaleryl-CoA. We also report the isolation and characterization of bktB (β-ketothiolase B), which encodes the BktB β-ketothiolase required for efficient production of PHBV in R. eutropha. 相似文献
45.
Protein evolution in different cellular environments: cytochrome b in sharks and mammals 总被引:4,自引:0,他引:4
DNA sequences for the mitochondrial cytochrome b gene were determined for
13 species of sharks. Rates and patterns of amino acid replacement are
compared for sharks and mammals. Absolute rates of cytochrome b evolution
are six times slower in sharks than in mammals. Bivariate plots of the
number of nonsynonymous and silent transversions are indistinguishable in
the two groups, however, suggesting that the differences in amino acid
replacement rates are due primarily to differences in DNA substitution
rates. Patterns of amino acid replacement are also similar in the two
groups. Conserved and variable regions occur in the same parts of the
cytochrome b gene, and there is little evidence that the types of amino
acid changes are significantly different between the groups. Similarity in
the relative rates and patterns of protein change between the two groups
prevails despite dramatic differences in the cellular environments of
sharks and mammals. Poor penetrance of physiological differences through to
rates of protein evolution provides support for the neutral theory and
suggests that, for cytochrome b, patterns of evolution have been relatively
constant throughout much of vertebrate history.
相似文献
46.
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48.
Site-directed mutagenesis of a conserved region of the 5-enolpyruvylshikimate-3-phosphate synthase active site 总被引:11,自引:0,他引:11
S R Padgette D B Re C S Gasser D A Eichholtz R B Frazier C M Hironaka E B Levine D M Shah R T Fraley G M Kishore 《The Journal of biological chemistry》1991,266(33):22364-22369
The active site of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) has been probed using site-directed mutagenesis and inhibitor binding techniques. Replacement of a specific glycyl with an alanyl or a prolyl with a seryl residue in a highly conserved region confers glyphosate tolerance to several bacterial and plant EPSPS enzymes, suggesting a high degree of structural conservation between these enzymes. The glycine to alanine substitution corresponding to Escherichia coli EPSPS G96A increases the Ki(app) (glyphosate) of petunia EPSPS 5000-fold while increasing the Km(app)(phosphoenolpyruvate) about 40-fold. Substitution of this glycine with serine, however, abolishes EPSPS activity but results in the elicitation of a novel EPSP hydrolase activity whereby EPSP is converted to shikimate 3-phosphate and pyruvate. This highly conserved region is critical for the interaction of the phosphate moiety of phosphoenolpyruvate with EPSPS. 相似文献
49.
50.
Esophageal cancer involves multiple genetic alternations. A systematic codon usage bias analysis was completed to investigate the bias among the esophageal cancer responsive genes. GC-rich genes were low (average effective number of codon value was 49.28). CAG and GTA are over-represented and under-represented codons, respectively. Correspondence analysis, neutrality plot, and parity rule 2 plot analysis confirmed the dominance over mutation pressure in modulating the codon usage pattern of genes linked with esophageal cancer. 相似文献