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1.
A simple new procedure was described for producing a sequential series of overlapping clones for use in DNA sequencing. The technique used single-stranded M13 DNA and complementary DNA oligomers to form specific cleavage and ligation substrates. It was, therefore, independent of the sequence of the DNA cloned into the vector. Deletions of varying sizes were generated from one end of the insert through the 3' to 5' exonuclease activity of T4 DNA polymerase. The approximate size of the deletion and therefore the starting point for DNA sequencing could be estimated by electrophoresis of the subcloned phage DNA on a agarose gel. This greatly reduced the number of templates that must be sequenced to obtain a complete sequence. The entire procedure could be carried out in one tube in less than a day. The procedure was used to subclone and sequence the maize mitochondrial 18 S rDNA and 5' flanking region (2622 bases) in less than a week. Other applications of oligomers and single-stranded DNA in the construction of insertions, deletions, and cDNAs are discussed. 相似文献
2.
C. L. McIntyre B. Winberg K. Houchins R. Appels B. R. Baum 《Plant Systematics and Evolution》1992,183(3-4):249-264
Relationships between 9Oryza species, covering 6 different genomes, have been studied using hybridization and nucleotide sequence information from the5S Dna locus. Four to five units of the major size class of 5S DNA in each species, 55 units in all, were cloned and sequenced. Both hybridization and sequence data confirmed the basic differences between the A and B, C, D genome species suggested by morphological and cytological data. The 5S DNA units of the A genome species were very similar, as were the ones from the B, C, and D genome-containing species. The 5S DNA ofO. australiensis (E genome) grouped with the B, C, D cluster, while the units ofO. brachyantha (F genome) were quite different and grouped away from all other species. 5S DNA units fromO. minuta, O. latifolia, O. australiensis, andO. brachyantha hybridized strongly, and preferentially, to the genomic DNA from which the units were isolated and hence could be useful as species/genome specific probes. The 5S DNA units fromO. sativa, O. nivara, andO. rufipogon provided A genome-specific probes as they hybridized preferentially to A genome DNA. The units fromO. punctata andO. officinalis displayed weaker preferential hybridization toO. punctata DNA, possibly reflecting their shared genome (C genome). 相似文献
3.
DNA sequence analysis of a 5.27-kb direct repeat occurring adjacent to the regions of S-episome homology in maize mitochondria. 总被引:8,自引:1,他引:7
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J P Houchins H Ginsburg M Rohrbaugh R M Dale C L Schardl T P Hodge D M Lonsdale 《The EMBO journal》1986,5(11):2781-2788
The DNA sequence of the 5270-bp repeated DNA element from the mitochondrial genome of the fertile cytoplasm of maize has been determined. The repeat is a major site of recombination within the mitochondrial genome and sequences related to the R1(S1) and R2(S2) linear episomes reside immediately adjacent to the repeat. The terminal inverted repeats of the R1 and R2 homologous sequences form one of the two boundaries of the repeat. Frame-shift mutations have introduced 11 translation termination codons into the transcribed S2/R2 URFI gene. The repeated sequence, though recombinantly active, appears to serve no biological function. 相似文献
4.
The effect of 2-(n-heptyl)-4-hydroxyquinoline N-oxide (HQNO) on the kinetics of cytochrome b-563 and cytochrome c2 turnovers following single-turnover flashes was measured in isolated heterocysts. Low concentrations of HQNO (below 3 μM) blocked reoxidation of cytochrome b-563, whereas higher concentrations (above 5 μM) resulted in additional inhibition of cytochrome b-563 oxidation and also inhibited reduction of cytochrome b-563 and cytochrome c. Similar effects on cytochrome b-563 reduction and reoxidation were obtained with a combination of 5 μM HQNO and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (1–7 μM). In HQNO-inhibited heterocysts, cytochrome c reduction following a flash occurred in three phases with half-times of 0.5, 2.8 and 45 ms. The second phase nearly equalled the cytochrome b-563 reduction in half-time and magnitude. In the presence of HQNO, the reoxidation of cytochrome b-563 following two closely spaced actinic flashes displayed biphasic kinetics. The two phases correspond to reoxidation of cytochrome b-563 in which one or both of the cytochrome b-563 hemes in the cytochrome b–f complex are reduced. These results are interpreted in terms of a Q-loop in which HQNO, at low concentrations, blocks the site of rapid cytochrome b-563 reoxidation and at higher concentrations, also inhibits the site of electron donation by plastoquinol to the cytochrome b-f complex. 相似文献
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DANIEL TRAN TAKASHI KADONO MARIA LIA MOLAS RAFIK ERRAKHI JOËL BRIAND BERNADETTE BILIGUI TOMONORI KAWANO FRANÇOIS BOUTEAU 《Plant, cell & environment》2013,36(3):569-578
Ozone (O3) is an air pollutant with an impact increasingly important in our industrialized world. It affects human health and productivity in various crops. We provide the evidences that treatment of Arabidopsis thaliana with O3 results in ascorbate‐derived oxalic acid production. Using cultured cells of A. thaliana as a model, here we further showed that oxalic acid induces activation of anion channels that trigger depolarization of the cell, increase in cytosolic Ca2+ concentration, generation of reactive oxygen species and cell death. We confirmed that O3 reacts with ascorbate in the culture, thus resulting in production of oxalic acid and this could be part of the O3‐induced signalling pathways that trigger programmed cell death. 相似文献
7.
Preliminary estimates of the potential for carbon mitigation in European soils through no-till farming 总被引:9,自引:0,他引:9
PETE. SMITH DAVID S. POWLSON MARGARET J. GLENDINING JO. U. SMITH 《Global Change Biology》1998,4(6):679-685
In this paper we estimate the European potential for carbon mitigation of no-till farming using results from European tillage experiments. Our calculations suggest some potential in terms of (a) reduced agricultural fossil fuel emissions, and (b) increased soil carbon sequestration. We estimate that 100% conversion to no-till farming would be likely to sequester about 23 Tg C y–1 in the European Union or about 43 Tg C y–1 in the wider Europe (excluding the former Soviet Union). In addition, up to 3.2 Tg C y–1 could be saved in agricultural fossil fuel emissions. Compared to estimates of the potential for carbon sequestration of other carbon mitigation options, no-till agriculture shows nearly twice the potential of scenarios whereby soils are amended with organic materials. Our calculations suggest that 100% conversion to no-till agriculture in Europe could mitigate all fossil fuel-carbon emissions from agriculture in Europe. However, this is equivalent to only about 4.1% of total anthropogenic CO2-carbon produced annually in Europe (excluding the former Soviet Union) which in turn is equivalent to about 0.8% of global annual anthropogenic CO2-carbon emissions. 相似文献
8.
ISABELLE DE CAUWER MATHILDE DUFAY JOËL CUGUEN JEAN‐FRANÇOIS ARNAUD 《Molecular ecology》2010,19(8):1540-1558
Plant mating systems are known to influence population genetic structure because pollen and seed dispersal are often spatially restricted. However, the reciprocal outcomes of population structure on the dynamics of polymorphic mating systems have received little attention. In gynodioecious sea beet (Beta vulgaris ssp. maritima), three sexual types co‐occur: females carrying a cytoplasmic male sterility (CMS) gene, hermaphrodites carrying a non‐CMS cytoplasm and restored hermaphrodites that carry CMS genes and nuclear restorer alleles. This study investigated the effects of fine‐scale genetic structure on male reproductive success of the two hermaphroditic forms. Our study population was strongly structured and characterized by contrasting local sex‐ratios. Pollen flow was constrained over short distances and depended on local plant density. Interestingly, restored hermaphrodites sired significantly more seedlings than non‐CMS hermaphrodites, despite the previous observation that the former produce pollen of lower quality than the latter. This result was explained by the higher frequency of females in the local vicinity of restored (CMS) hermaphrodites as compared to non‐CMS hermaphrodites. Population structure thus strongly influences individual fitness and may locally counteract the expected effects of selection, suggesting that understanding fine scale population processes is central to predicting the evolution of gender polymorphism in angiosperms. 相似文献
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10.
Rye Jin KIM Seung Han OH Mi Young NOH Yong Hun JO Seongon KIM In Seok BANG Yong Seok LEE Iksoo KIM Yeon Soo HAN 《Entomological Research》2010,40(4):217-224
Calreticulin (CRT) plays pivotal roles in Ca2+ homeostasis, molecular chaperoning, infection, inflammation and innate immunity. In an attempt to study the involvement of CRT in innate immunity, the full-length cDNA of calreticulin (PxCRT) was cloned from the diamondback moth, Plutella xylostella. It consists of 1674 bp (excluding poly-A tail) with a longest open reading frame (ORF) of 1197 bp encoding 398 amino acids. In silico analysis of PxCRT ORF reveals that it has various repeat motifs and endoplasmic reticulum retention signal found in all the calreticulin proteins. As expected, high amino acid sequence identities were found from other CRTs identified from Bombyx mori (87%), Galleria mellonella (87%), Apis mellifera (74%), Anopheles gambiae (74%), Tribolium castaneum (73%), Culex quinquefasciatus (73%), Rhodnius prolixus (72%), Nasonia vitripennis (71%), Drosophila melanogaster (71%) and Haemaphysalis qinghaiensis (68%). During development, P. xylostella expressed PxCRT predominantly in the pupal stage. In addition, spatial expression pattern analysis indicates that PxCRT was highly expressed in the silk gland. PxCRT mRNA, furthermore, was strongly induced 3 to 6 h after laminarin treatment, suggesting that PxCRT appears to be involved in immune responses and also plays an important role in the silk gland. 相似文献