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  • 1
    ISSN: 0378-1119
    Keywords: Gene cloning ; PCR ; genome analysis ; satellite DNA ; stability analysis ; sugar beet
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0370-2693
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Physics
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1041
    Keywords: anxiety ; atenolol ; β-blockers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary In a randomized double-blind prospective between-patients trial in patients presenting with primary anxiety, atenolol significantly improved mean values on the Hamilton rating scale at two and four weeks when compared with placebo. There was also a significant improvement in affective symptoms at 28 days for atenolol when compared with placebo.
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  • 4
    ISSN: 1432-2242
    Keywords: Key words Simple sequence repeats ; Linkage maps ; Sugar beet ; Marker-assisted selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A set of informative simple sequence repeat markers has been identified for use in the marker-assisted breeding of Beta vulgaris. Highly enriched small insert genomic libraries were constructed, consisting of 1536 clones (with inserts of between 250–900 bp). Screening the clones with CA, CT, CAA, CATA and GATA nucleotide-repeat probes revealed positive hybridisation to over 50% of the clones. Of these 340 were sequenced. Primer pairs were designed for sequences flanking the repeats and, of these, 57 pairs revealed length polymorphism with 12 Beta accessions. Heterozygosity levels of the SSR loci ranged from 0.069 to 0.809. Heterozygosity levels were found to be similar to those detected employing RFLP probes with the same accessions. Phenetic analysis using the markers, indicated relationships in accordance with known pedigrees. Twenty three of the SSR markers were polymorphic in one or both of two F2 mapping populations, and were placed relative to a framework of RFLP probes. The markers are distributed over all nine linkage groups of sugar beet.
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  • 5
    ISSN: 1432-2242
    Keywords: Key words Avicennia marina ; Microsatellite ; Mangrove ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An enriched microsatellite library of the mangrove species Avicennia marina was constructed, in which 85.8% of the clones contained microsatellite sequences. Of the microsatellite repeat sequences isolated, 55.0% were di-nucleotides, 34.2% were tri-nucleotides, 50.0% were perfect, 24.2% were imperfect, and 15.0% were compound. Four different di-nucleotide repeats were isolated with repeat lengths ranging from 5 to 33; ten different tri-nucleotide repeats were isolated with repeat lengths ranging from 3 to 25. The most common di-nucleotide was the AC/TG repeat; the most common tri-nucleotide was the CCG/GGC repeat. Sixteen microsatellite sequences were selected for primer design, and 6 primers were selected to investigate the polymorphism detected among 15 individuals of A. marina from three natural populations in Australia. A total of 40 alleles were detected at 6 microsatellite loci. The number of alleles per microsatellite locus ranged from 5 to 13. On average, 7 alleles were detected per locus. All microsatellite loci showed high levels of gene diversity (heterozygosity), with values ranging from 0.53 to 0.88; the mean value of gene diversity was 0.70. Microsatellite loci were also tested for conservation across Avicennia species. There was a decline in amplification success with increasing divergence between Avicennia species. The results indicate that microsatellites are abundant in the Avicennia genome and can be valuable genetic markers for assessing the effects of deforestation and forest fragmentation in mangrove communities, which is an important issue for mangrove conservation and afforestation schemes.
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  • 6
    ISSN: 1432-2242
    Keywords: Key words Simple sequence repeats ; Inter-specific priming ; Molecular markers ; Grapevine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Eleven microsatellites isolated from grapevine (Vitis vinifera) were used to study the degree of conservation of these sequences across different Vitis species. Nine microsatellites were newly isolated, the remaining two (VVS2 and VVS5) came from the literature. A preliminary assay on the conservation of priming sites was carried out on 14 non-V. vinifera species, including relevant taxa for breeding. Parthenocissus quinquefolia was added as representative of a related genus. Cross-species amplification was obtained in 94% of the 176 genotype×locus tested combinations. Three microsatellite loci were then cloned and sequenced in ten species. The microsatellite repeat was found present in all cases. The repeat region was often longer in V. vinifera than in the other species. Furthermore the non-source species showed interruptions in the repeat. In spite of these constraints, which could reduce the polymorphism of microsatellites in non-source species, the results demonstrate the possibility of extending the use of microsatellite markers to wild germplasm and inter-specific hybrids. Point mutations have been found in microsatellite flanking regions and these variations have been used to investigate the genetic relationship among taxa. The Neighbor-joining tree that was obtained on the basis of ten nucleotide variations, showed that there is not a clear cut difference between American, Asian and European species and that the actual taxonomy which reflects the geographical distribution of species must most likely be revised. Moreover, in general, nucleotide variations which occur in microsatellite flanking regions provide new molecular tools for investigating the evolution of species.
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  • 7
    ISSN: 1432-2242
    Keywords: Key words Bread wheat ; Grain protein content ; Microsatellite ; STMS ; QTL analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  This study was undertaken with a view to tag gene(s) controlling grain protein content (GPC) using molecular markers in bread wheat. For this purpose, the genotype PH132 with high protein content (13.5%) was crossed with genotype WL711 with significantly lower protein content (9.7%), and 100 RILs were derived. These RILs showed normal distribution for protein content. The parental genotypes were analysed with 232 STMS primer pairs for detection of polymorphism. Of these, 167 primer pairs gave scorable amplification products, and 57 detected polymorphism between the parents. Using each of these 57 primer pairs, we carried out bulked segregant analysis on RILs representing the two extremes of the distribution. One primer pair for the locus wmc41 showed association with protein content. This was further confirmed through selective genotyping. The co-segregation data on the molecular marker (wmc41) and protein content on 100 RILs was analysed by means of a single-marker linear regression approach. Significant regression suggested linkage between wmc41 and a QTL (designated as QGpc.ccsu-2D.) for protein content. The results showed that this marker-linked QTL accounted for 18.73% of the variation for protein content between the parents. The marker has been located on chromosome arm 2DL using nulli-tetrasomic lines and two ditelocentric stocks for chromosome 2D.
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  • 8
    ISSN: 1432-2242
    Keywords: Key words Preharvest sprouting ; Microsatellite ; STMS ; STS ; Linkage ; Bread wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In bread wheat, the transfer of tolerance to preharvest sprouting (PHS) that is associated with genotypes having red kernel colour to genotypes with amber kernels is difficult using conventional methods of plant breeding. The study here was undertaken to identify DNA markers linked with tolerance to PHS as these would allow indirect marker-assisted selection of PHS-tolerant genotypes with amber kernels. For this purpose, a set of 100 recombinant inbred lines (RILs) was developed using a cross between a PHS-tolerant genotype, SPR8198, with red kernels and a PHS-susceptible cultivar, ‘HD2329’, with white kernels. The two parents were analysed with 232 STMS (sequence-tagged microsatellite site) and 138 STS (sequence-tagged site) primer pairs. A total of 300 (167 STMSs and 133 STSs) primer pairs proved functional by giving scorable PCR products. Of these, 57 (34%) STMS and 30 (23%) STS primer pairs detected reproducible polymorphism between the parent genotypes. Using these primer pairs, we carried out bulked segregant analysis on two bulked DNAs, one obtained by pooling DNA from 5 PHS-tolerant RILs and the other similarly derived by pooling DNA from 5 PHS-susceptible RILs. Two molecular markers, 1 STMS primer pair for the locus wmc104 anda STS primer pair for the locus MST101, showed apparent linkage with tolerance to PHS. This was confirmed following selective genotyping of individual RILs included in the bulks. Chi-square contingency tests for independence were conducted on the cosegregation data collected on 100 RILs involving each of the two molecular markers (wmc104 and MST101) and PHS. The tests revealed a strong association between each of the markers and tolerance to PHS. Using nullisomic-tetrasomic lines, we were able to assign wmc104 and MST101 to chromosomes 6B and 7D, respectively. The results also indicated that the tolerance to PHS in SPR8198 is perhaps governed by two genes (linked with two molecular markers) exhibiting complementary interaction.
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  • 9
    ISSN: 1432-2242
    Keywords: Hexaploid wheat ; Endosperm ; Allelic variation ; α-Amylase ; Isozymes ; Isoelectric focusing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A study of α-amylase isozyme patterns from gibberellin-induced endosperms from more than 200 wheat genotypes has revealed allelic variation at five of the six α-Amy-1 and α-Amy-2 structural loci. These differences will find application as genetic markers and in varietal identification. The α-Amy-B1 locus on chromosome 6B was most variable and displayed eight distinct allelic forms. The nature of the allelic phenotypes, observations of segregating populations and the number of in vivo translation products of mRNAs from the α-Amy-1 and α-Amy-2 loci indicated that the individual loci are multigenic, each consisting of tightly linked subunits which produce several different isoforms.
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  • 10
    ISSN: 1432-2242
    Keywords: Beta vulgaris ; Polymerase chain reaction ; Yeast artificial chromosome library ; Genome mapping ; Restriction fragment length polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A yeast artificial chromosome (YAC) genomic library of Beta vulgaris was constructed in the pYAC4 vector. High-molecular-weight DNA was prepared from agarose-embedded leaf protoplasts from a triploid cultivar. The library was found to contain 33,500 clones in an ordered array of microtiter plates. Mean size of the inserts was estimated to be 135 kb, and the library should therefore represent the equivalent of five haploid genomes. The library was characterised for the presence of highly repetitive, chloroplast and single-copy sequences. In order to isolate single-copy sequences, 18 pools of DNA, each from 1920 individual YAC clones, were prepared for rapid screening of the library by the polymerase chain reaction. The results of these screenings showed that the number of isolated clones was at or near the frequency expected.
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