scholarly journals Detection of Single-Copy Sequences with Digoxigenin-Labeled Probes in a Complex Plant Genome After Separation on Pulsed-Field Gels

BioTechniques ◽  
1996 ◽  
Vol 21 (6) ◽  
pp. 1067-1072 ◽  
Author(s):  
T. Lahaye ◽  
B. Rueger ◽  
S. Toepsch ◽  
J. Thalhammer ◽  
P. Schulze-Lefert
Keyword(s):  
Single Copy ◽  
Plant Genome ◽  
Pulsed Field

Construction of a MluI-YAC library from barley (Hordeum vulgare L.) and analysis of YAC insert terminal regions

Genome ◽  
1997 ◽  
Vol 40 (6) ◽  
pp. 896-902 ◽  
Author(s):  
Michael Kleine ◽  
Christian Jung ◽  
Wolfgang Michalek ◽  
Thomas Diefenthal ◽  
Harald Dargatz
Keyword(s):  
Hordeum Vulgare ◽  
Gel Electrophoresis ◽  
Yeast Artificial Chromosome ◽  
Single Copy ◽  
Pulsed Field Gel Electrophoresis ◽  
Hordeum Vulgare L ◽  
High Molecular Weight Dna ◽  
Pulsed Field ◽  
Artificial Chromosomes ◽  
Yac Library

We describe the construction of a specific yeast artificial chromosome (YAC) library from barley (Hordeum vulgare L.) using the vector pYAC-RC. The library was generated by total digestion of high molecular weight DNA with the infrequently cutting restriction enzyme MluI. Only 10–30% of the colonies were recombinant, as visualized by red–white selection and subsequent pulsed-field gel electrophoresis analysis. About 17 000 individual recombinant YAC clones with insert sizes ranging from 50 to 700 kb, with a mean of 170 kb, were selected. No chloroplast sequences were detected and the proportion of YAC clones containing BARE–1 copia–like retroelements is about 5%. Screening of the library with a single-copy RFLP marker closely linked to the Mla locus yielded three identical clones of the same size. Insert termini of randomly chosen YAC clones were investigated with respect to their redundancy in the barley genome and compared with termini of YAC clones from an EcoRI-based YAC library, resulting in a fourfold enrichment of single-copy sequences at the MluI vector–insert junctions.Key words: yeast artificial chromosomes, YAC, Hordeum vulgare, pulsed-field gel electrophoresis.

Nonisotopic in situ hybridization and plant genome mapping: the first 10 years

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 717-725 ◽  
Author(s):  
Jiming Jiang ◽  
Bikram S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Physical Mapping ◽  
Dna Sequences ◽  
Genome Mapping ◽  
Molecular Cytogenetics ◽  
Gene Families ◽  
Single Copy ◽  
Plant Genome ◽  
Metaphase Chromosomes

Nonisotopic in situ hybridization (ISH) was introduced in plants in 1985. Since then the technique has been widely used in various areas of plant genome mapping. ISH has become a routine method for physical mapping of repetitive DNA sequences and multicopy gene families. ISH patterns on somatic metaphase chromosomes using tandemly repeated sequences provide excellent physical markers for chromosome identification. Detection of low or single copy sequences were also reported. Genomic in situ hybridization (GISH) was successfully used to analyze the chromosome structure and evolution of allopolyploid species. GISH also provides a powerful technique for monitoring chromatin introgession during interspecific hybridization. A sequential chromosome banding and ISH technique was developed. The sequential technique is very useful for more precise and efficient mapping as well as cytogenetic determination of genomic affinities of individual chromosomes in allopolyploid species. A critical review is made on the present resolution of the ISH technique and the future outlook of ISH research is discussed.Key words: in situ hybridization, physical mapping, genome mapping, molecular cytogenetics.

Expression of the DREB1A gene in lentil (Lens culinaris Medik. subsp. culinaris) transformed with the Agrobacterium system

2011 ◽  
Vol 62 (6) ◽  
pp. 488 ◽  
Author(s):  
Fateh Khatib ◽  
Antonios Makris ◽  
Kasuko Yamaguchi-Shinozaki ◽  
Shiv Kumar ◽  
Ashtuosh Sarker ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Lens Culinaris ◽  
Marker Gene ◽  
Single Copy ◽  
Plant Genome ◽  
Direct Organogenesis ◽  
Complete Resistance ◽  
Protocol Optimisation ◽  
Gusa Gene ◽  
Rd29a Promoter

Until now three publications have reported the development of transgenic lentil plants through protocol optimisation using the gusA gene, but there are no reports of the introduction of a gene with agronomic importance. In the present study we report the introduction of the DREB1A gene into lentil to enhance drought and salinity tolerance. Decapitated embryos were immersed in Agrobacterium suspension and then co-cultivated for 4 days. Direct organogenesis was induced from the apical meristems and cotyledonary buds. Subsequently, the explants were subjected to selection in medium containing 10 mg/L phosphinothricin for nine rounds with 2-week intervals. The putative transgenic explants were micro-grafted onto non-transformed rootstocks to establish transgenic plants. The PCR results confirmed the insertion and stable inheritance of the gene of interest and bar marker gene in the plant genome. The Southern blot analysis revealed the integration of a single copy of the transgenes. T0 plants and progeny up to T2 generations showed complete resistance to the herbicide Basta. The DREB1A gene driven by the rd29A promoter was induced in transgenic plants by salt stress from sodium chloride solution. The total RNA was extracted and cDNA synthesised. The results showed that DREB1A mRNA was accumulated and thus the DREB1A transgene was expressed in the transgenic plants, whereas no expression was detected in the non-transformed parents.

scholarly journals Two ancient genome duplication events shape diversity in Hibiscus L. (Malvaceae)

2020 ◽  
Author(s):  
Jonna Sofia Eriksson ◽  
Christine D. Bacon ◽  
Dominic J. Bennett ◽  
Bernard E. Pfeil ◽  
Bengt Oxelman ◽  
...  
Keyword(s):  
Chromosome Number ◽  
Genome Size ◽  
Genome Duplication ◽  
Single Copy ◽  
Genomic Diversity ◽  
Plant Genome ◽  
Genome Doubling ◽  
Gene Copies ◽  
Plant Groups ◽  
Duplication Events

Abstract Background: The great diversity in plant genome size and chromosome number is partly due to polyploidization (i.e., genome doubling events). The differences in genome size and chromosome number among diploid plant species can be a window into the intriguing phenomenon of past genome doubling that may be obscured through time by the process of diploidization. The genus Hibiscus L. (Malvaceae) has a wide diversity of chromosome numbers and an allegedly complex genomic history. Hibiscus is ideal for exploring past genomic events because although two ancient genome duplication events have been identified, as more are still likely to be found, considering its diverse background. To reappraise the group’s genomic history, we tested a series of scenarios describing different polyploidization events using previously identified duplications in Hibiscus syriacus.Results: The data showed that >54% of the single-copy genes where in fact paralogues. When testing for different genome duplication scenarios using gene count data; species of Hibiscus was shown to have shared one genome duplication with H. syriacus, -- while one whole genome duplication was contained within H. syriacus, -- to be the preferred model given the observed distribution of paralogous gene copies in Hibiscus.Conclusions: Here, we corroborated the independent genome doubling previously found in the lineage leading to H. syriacus and a shared genome doubling of this lineage and the remainder of Hibiscus. Additionally, we found a previously undiscovered genome duplication shared by the /Pavonia and /Malvaviscus clades (both nested within Hibiscus) with the occurrences of two copies in what were otherwise single-copy genes. Our results highlight the complexity of genomic diversity in some plant groups, which makes orthology assessment and accurate phylogenomic inference difficult.

Identification of a single-copy gene encoding a Type I chlorophyll a/b-binding polypeptide of photosystem I in Arabidopsis thaliana

1992 ◽  
Vol 84 (4) ◽  
pp. 561-567 ◽  
Author(s):  
Poul E. Jensen ◽  
Michael Kristensen ◽  
Tine Hoff ◽  
Jan Lehmbeck ◽  
Bjarne M. Stummann ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Chlorophyll A ◽  
Photosystem I ◽  
Single Copy ◽  
Type I ◽  
Single Copy Gene ◽  
Gene Encoding ◽  
Copy Gene

ADSORPTION OF NO ON Pt STUDIED BY PULSED FIELD DESORPTION MASS SPECTROMETRY

1984 ◽  
Vol 45 (C9) ◽  
pp. C9-227-C9-230
Author(s):  
N. Kruse ◽  
T. Kessler ◽  
G. Abend ◽  
J. H. Block
Keyword(s):  
Mass Spectrometry ◽  
Field Desorption ◽  
Desorption Mass Spectrometry ◽  
Pulsed Field ◽  
Field Desorption Mass Spectrometry

Controlling Gas Selectivity in Molecular Porous Liquids by Tuning the Cage Window Size

2019 ◽  
Author(s):  
Benjamin Egleston ◽  
Konstantin V. Luzyanin ◽  
Michael C. Brand ◽  
Rob Clowes ◽  
Michael E. Briggs ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Chemical Synthesis ◽  
Field Gradient ◽  
Window Size ◽  
Standard Approach ◽  
Porous Solids ◽  
Pulsed Field Gradient Nmr ◽  
Pulsed Field ◽  
Cage Molecules ◽  
Gas Selectivity

Control of pore window size is the standard approach for tuning gas selectivity in porous solids. Here, we present the first example where this is translated into a molecular porous liquid formed from organic cage molecules. Reduction of the cage window size by chemical synthesis switches the selectivity from Xe-selective to CH<sub>4</sub>-selective, which is understood using <sup>129</sup>Xe, <sup>1</sup>H, and pulsed-field gradient NMR spectroscopy.

Controlling Gas Selectivity in Molecular Porous Liquids by Tuning the Cage Window Size

2019 ◽  
Author(s):  
Benjamin Egleston ◽  
Konstantin V. Luzyanin ◽  
Michael C. Brand ◽  
Rob Clowes ◽  
Michael E. Briggs ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Chemical Synthesis ◽  
Field Gradient ◽  
Window Size ◽  
Standard Approach ◽  
Porous Solids ◽  
Pulsed Field Gradient Nmr ◽  
Pulsed Field ◽  
Cage Molecules ◽  
Gas Selectivity

Control of pore window size is the standard approach for tuning gas selectivity in porous solids. Here, we present the first example where this is translated into a molecular porous liquid formed from organic cage molecules. Reduction of the cage window size by chemical synthesis switches the selectivity from Xe-selective to CH<sub>4</sub>-selective, which is understood using <sup>129</sup>Xe, <sup>1</sup>H, and pulsed-field gradient NMR spectroscopy.

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