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91.
Intraspecific DNA sequence variation of the mitochondrial control region of white sturgeon (Acipenser transmontanus) 总被引:3,自引:0,他引:3
Intraspecific sequence variation in the D-loop region of mtDNA in white
sturgeon (Acipenser transmontanus), a relict North American fish species,
was examined in 27 individuals from populations of the Columbia and Fraser
rivers. Thirty-three varied nucleotide positions were present in a
462-nucleotide D-loop sequence, amplified using the polymerase chain
reaction. Bootstrapped neighbor-joining and maximum- parsimony trees of
sequences from 19 haplotypes suggest that the two populations have recently
diverged. This is consistent with the hypothesis that the Columbia River, a
Pleistocene refugium habitat, was the source of founders for the Fraser
River after the last glacial recession. On the basis of a divergence time
of 10-12 thousand years ago, the estimated substitution rate of the white
sturgeon D-loop region is 1.1-1.3 x 10(-7) nucleotides/site/year, which is
comparable to rates for hypervariable sequences in the human D-loop region.
Furthermore, the ratio of mean percent nucleotide differences in the D-
loop (2.27%) to that in whole mtDNA (0.54%, as estimated from
restriction-enzyme data) is 4.3, which is similar to the fourfold-to-
fivefold-higher substitution rate estimated for the human D-loop. The high
nucleotide substitution rate of the hypervariable region indicates that the
vertebrate D-loop has potential as a genetic marker in molecular population
studies.
相似文献
92.
Two mutations in the HMG-box with very different structural consequences provide insights into the nature of binding to four-way junction DNA. 总被引:4,自引:3,他引:1 下载免费PDF全文
S H Teo K D Grasser C H Hardman R W Broadhurst E D Laue J O Thomas 《The EMBO journal》1995,14(15):3844-3853
Mutation of the highly conserved tryptophan residue in the A-domain HMG-box of HMG1 largely, but not completely, destroys the protein tertiary structure and abolishes its supercoiling ability, but does not abolish structure-specific DNA binding to four-way junctions. Circular dichroism shows that the protein has some residual alpha-helix (< 10%) and does not re-fold in the presence of DNA. Structure-specific DNA binding might therefore be a property of some primary structure element, for example the N-terminal extended strand, which even in the unfolded protein would be held in a restricted conformation by two, largely trans, X-Pro peptide bonds. However, mutation of P5 or P8 of the A-domain to alanine does not abolish the formation of the (first) complex in a gel retardation assay, which probably arises from binding to the junction cross-over, although the P8 mutation does affect the formation of higher complexes which may arise from binding to the junction arms. Since mutation of P8 in the W49R mutant has no effect on structure-specific junction binding, we propose that some residual alpha-helix in the protein might be involved, implicating this element in the interactions of HMG-boxes generally with DNA. 相似文献
93.
Ancient large-scale genome duplications: phylogenetic and linkage analyses shed light on chordate genome evolution 总被引:12,自引:4,他引:8
Pebusque MJ; Coulier F; Birnbaum D; Pontarotti P 《Molecular biology and evolution》1998,15(9):1145-1159
Paralogous genes from several families were found in four human chromosome
regions (4p16, 5q33-35, 8p12-21, and 10q24-26), suggesting that their
common ancestral region underwent several rounds of large- scale
duplication. Searches in the EMBL databases, followed by phylogenetic
analyses, showed that cognates (orthologs) of human duplicated genes can be
found in other vertebrates, including bony fishes. In contrast, within each
family, only one gene showing the same high degree of similarity with all
the duplicated mammalian genes was found in nonvertebrates (echinoderms,
insects, nematodes). This indicates that large-scale duplications occurred
after the echinoderms/chordates split and before the bony vertebrate
radiation. It has been suggested that two rounds of gene duplication
occurred in the vertebrate lineage after the separation of Amphioxus and
craniate (vertebrates + Myxini) ancestors. Before these duplications, the
genes that have led to the families of paralogous genes in vertebrates must
have been physically linked in the craniate ancestor. Linkage of some of
these genes can be found in the Drosophila melanogaster and Caenorhabditis
elegans genomes, suggesting that they were linked in the triploblast
Metazoa ancestor.
相似文献
94.
Endogenous chloride channels of insect sf9 cells. Evidence for coordinated activity of small elementary channel units 总被引:1,自引:0,他引:1 下载免费PDF全文
EH Larsen SE Gabriei MJ Stutts J Fullton EM Price RC Boucher 《The Journal of general physiology》1996,107(6):695-714
The endogenous Cl- conductance of Spodoptera frugiperda (Sf9) cells was studied 20-35 h after plating out of either uninfected cells or cells infected by a baculovirus vector carrying the cloned beta-galactosidase gene (beta-Gal cells). With the cation Tris+ in the pipette and Na+ in the bath, the reversal potential of whole-cell currents was governed by the prevailing Cl- equilibrium potential and could be fitted by the Goldman-Hodgkin-Katz equation with similar permeabilities for uninfected and beta-Gal cells. In the frequency range 0.12 < f < 300 Hz, the power density spectrum of whole-cell Cl- currents could be fitted by three Lorentzians. Independent of membrane potential, >50% of the total variance of whole-cell current fluctuations was accounted for by the low frequency Lorentzian (fc = 0.40 +/- 0.03 Hz, n = 6). Single-Cl- channels showed complex gating kinetics with long lasting (seconds) openings interrupted by similar long closures. In the open state, channels exhibited fast burst-like closures. Since the patches normally contained more than a single channel, it was not possible to measure open and closed dwell-time distributions for comparing single-Cl- channel activity with the kinetic features of whole-cell currents. However, the power density spectrum of Cl- currents of cell-attached and excised outside-out patches contained both high and low frequency Lorentzian components, with the corner frequency of the slow component (fc = 0.40 +/- 0.02 Hz, n = 4) similar to that of whole-cell current fluctuations. Chloride channels exhibited multiple conductance states with similar Goldman-Hodgkin-Katz-type rectification. Single-channel permeabilities covered the range from approximately 0.6.10(-14) cm5/s to approximately 6.10(-14) cm3/s, corresponding to a limiting conductance (gamma 150/150) of approximately 3.5 pS and approximately 35 pS, respectively. All states reversed near the same membrane potential, and they exhibited similar halide ion selectivity, P1 > PCl approximately PBr. Accordingly, Cl- current amplitudes larger than current flow through the smallest channel unit resolved seem to result from simultaneous open/shut events of two or more channel units. 相似文献
95.
In vivo and in vitro processing of seed reserve protein in the endoplasmic reticulum: evidence for two glycosylation steps 总被引:12,自引:0,他引:12 下载免费PDF全文
Cotyledons of the common bean (Phaseolus vulgaris L.) synthesize large amounts of the reserve protein phaseolin. The polypeptides are synthesized on membrane-bound polysomes, pass through the endoplasmic reticulum (ER) and accumulate in protein bodies. For a study of the biosynthesis and processing of phaseolin, developing cotyledons were labeled with radioactive amino acids, glucosamine and mannose, and isolated fractions (polysomal RNA, polysomes, and rough ER) were used for in vitro protein synthesis. Newly synthesized phaseolin present in the ER of developing cotyledons can be fractioned into four glycopolypeptides by SDS PAGE. In vitro synthesis with polysomal RNA results in the formation of two polypeptides by polysome run-off shows that glycosylation is a co-translational event. The two unglycosylated polypeptides formed by polysome run-off are slightly smaller than the two polypeptides formed by in vitro translation of isolated RNA, indicating that a signal peptide may be present on these polypeptides. Run-off synthesis with rough ER produces a pattern of four polypeptides similar to the one obtained by in vivo labeling. The two abundant glycopolypeptides formed by polysome run-off. This result indicates the existence of a second glycosylation event for the abundant polypeptides. Inhibition of glycosylation by Triton X-100 during chain-completion with rough ER was used to show that these two glycosylation steps normally occur sequentially. Both glycosylation steps are inhibited by tunicamycin. Analysis of carhohydrate to protein ratios of the different polypeptides and of trypsin digests of polypeptides labeled with [(3)H]glucosamine confirmed the conclusion that some glycosylated polypeptides contain two oligosaccharide chains, while others contain only one. An analysis of tryptic peptide maps shows that each of the unglycosylated polypeptides is the precursor for one glycosylated polypeptide with one oligosaccharide chain and one with two oligosaccharide chains. 相似文献
96.
Substructure of the glomerular slit diaphragm in freeze-fractured normal rat kidney 总被引:4,自引:3,他引:1 下载免费PDF全文
In the renal glomerulus, the narrow slits between adjacent epithelial podocytes are bridged by a diaphragm (2, 8, 11). In rat and mouse kidneys fixed by perfusion with tannic acid and glutaraldehyde (TAG), it has recently been discovered that this diaphragm has a highly ordered, isoporous substructure (9). It consists of a regular array of alternating cross bridges extending from the podocyte plasma membranes to a centrally running filament. This zipperlike pattern results in two rows of rectangular pores, approximately 40 X 140 A in cross section, dimensions consistent with the proposed role of the diaphragm as an important filtration barrier to plasma proteins (6). In the present study, we found in freeze-cleaved and in freeze-etched normal rat glomeruli that the surface of the slit diaphragm has an appearance conforming to the pattern found in sectioned material. 相似文献
97.
H?Bukulmez AL?Matthews CM?Sullivan C?Chen MJ?Kraay RC?Elston RW?Moskowitz VM?Goldberg ML?WarmanEmail author 《Arthritis research & therapy》2005,8(1):R25
In order to determine whether there is a genetic component to hip or knee joint failure due to idiopathic osteoarthritis (OA),
we invited patients (probands) undergoing hip or knee arthroplasty for management of idiopathic OA to provide detailed family
histories regarding the prevalence of idiopathic OA requiring joint replacement in their siblings. We also invited their spouses
to provide detailed family histories about their siblings to serve as a control group. In the probands, we confirmed the diagnosis
of idiopathic OA using American College of Rheumatology criteria. The cohorts included the siblings of 635 probands undergoing
total hip replacement, the siblings of 486 probands undergoing total knee replacement, and the siblings of 787 spouses. We
compared the prevalence of arthroplasty for idiopathic OA among the siblings of the probands with that among the siblings
of the spouses, and we used logistic regression to identify independent risk factors for hip and knee arthroplasty in the
siblings. Familial aggregation for hip arthroplasty, but not for knee arthroplasty, was observed after controlling for age
and sex, suggesting a genetic contribution to end-stage hip OA but not to end-stage knee OA. We conclude that attempts to
identify genes that predispose to idiopathic OA resulting in joint failure are more likely to be successful in patients with
hip OA than in those with knee OA. 相似文献
98.
Discrimination of modes of action of antifungal substances by use of metabolic footprinting 总被引:4,自引:0,他引:4
Allen J Davey HM Broadhurst D Rowland JJ Oliver SG Kell DB 《Applied and environmental microbiology》2004,70(10):6157-6165
Diploid cells of Saccharomyces cerevisiae were grown under controlled conditions with a Bioscreen instrument, which permitted the essentially continuous registration of their growth via optical density measurements. Some cultures were exposed to concentrations of a number of antifungal substances with different targets or modes of action (sterol biosynthesis, respiratory chain, amino acid synthesis, and the uncoupler). Culture supernatants were taken and analyzed for their "metabolic footprints" by using direct-injection mass spectrometry. Discriminant function analysis and hierarchical cluster analysis allowed these antifungal compounds to be distinguished and classified according to their modes of action. Genetic programming, a rule-evolving machine learning strategy, allowed respiratory inhibitors to be discriminated from others by using just two masses. Metabolic footprinting thus represents a rapid, convenient, and information-rich method for classifying the modes of action of antifungal substances. 相似文献
99.
A (15)N-labelled peripheral-subunit binding domain (PSBD) of the dihydrolipoyl acetyltransferase (E2p) and the dimer of a solubilized interface domain (E3int) derived from the dihydrolipoyl dehydrogenase (E3) were used to investigate the basis of the interaction of E2p with E3 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus. Thirteen of the 55 amino acids in the PSBD show significant changes in either or both of the (15)N and (1)H amide chemical shifts when the PSBD forms a 1 : 1 complex with E3int. All of the 13 amino acids reside near the N-terminus of helix I of PSBD or in the loop region between helix II and helix III. (15)N backbone dynamics experiments on PSBD indicate that the structured region extends from Val129 to Ala168, with limited structure present in residues Asn126 to Arg128. The presence of structure in the region before helix I was confirmed by a refinement of the NMR structure of uncomplexed PSBD. Comparison of the crystal structure of the PSBD bound to E3 with the solution structure of uncomplexed PSBD described here indicates that the PSBD undergoes almost no conformational change upon binding to E3. These studies exemplify and validate the novel use of a solubilized, truncated protein domain in overcoming the limitations of high molecular mass on NMR spectroscopy. 相似文献
100.
Restoring grassy woodland diversity through direct seeding: Insights from six ‘best‐practice’ case studies in southern Australia 下载免费PDF全文
Peter Cuneo Paul Gibson‐Roy Graham Fifield Linda Broadhurst Tim Berryman Andrew Crawford David Freudenberger 《Ecological Management & Restoration》2018,19(2):124-135
Ecological restoration of grassy woodland ecosystems is now a significant landscape‐scale conservation objective throughout southern Australia. Technological improvements in direct seeding are now sufficiently well‐advanced to examine whether cost‐effective restoration of grassy woodlands is feasible. Consideration of six ‘best practice case studies shows substantial evidence of success. Further refinement of direct seeding techniques, in combination with native seed production systems, however, will be required into the future to meet the scale of woodland conservation targets and restore ecological function. 相似文献