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Isolation of matrices from maize leaf nuclei: identification of a matrix-binding site adjacent to theAdh1 gene (1993)

Avramova, Zoya ; Bennetzen, Jeffrey L.

Springer

Plant molecular biology 22 (1993), S. 1135-1143

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ISSN: 1573-5028

Keywords: maize ; Adh1 gene ; nuclear matrix ; MAR

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract Nuclear matrices were isolated from maize leaves by the two conventional methods usually employed for the preparation of the corresponding structures of animal origin. It is demonstrated that functionally competent matrices, recognizing and specifically binding the MAR-containing DNA of the mousek-immunoglobulin gene may be prepared by both 2 M NaCl and LIS extractions of maize nuclei. A DNA region with a high affinity for the nuclear matrix was identified at the 5′ end of the maizeAdh1-S gene, distal to the promoter region. The presence of sites of reported altered chromatin structure in this particular region is discussed. While the proximity and the cohabitation of MARs with different regulatory elements is a common feature of matrix association regions in animal systems, this is the first plant MAR identified in a region of known significance for gene regulation.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00028982

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Recombination at the Rp1 locus of maize (1991)

Hulbert, Scot H. ; Bennetzen, Jeffrey L.

Springer

Molecular genetics and genomics 226 (1991), S. 377-382

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ISSN: 1617-4623

Keywords: Disease resistance ; Maize ; Recombination ; Unequal crossing-over ; Fine structure mapping

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Summary The Rp1 locus of maize determines resistance to races of the maize rust fungus (Puccinia sorghi). Restriction fragment length polymorphism markers that closely flank Rp1 were mapped and used to study the genetic fine structure and role of recombination in the instability of this locus. Susceptible progeny, lacking the resistance of either parent, were obtained from test cross progeny of several Rp1 heterozygotes. These susceptible progeny usually had non-parental genotypes at flanking marker loci, thereby verifying their recombinational origin. Seven of eight Rp1 alleles (or genes) studied were clustered within about 0.2 map units of each other. Rpl G, however, mapped from 1–3 map units distal to other Rp1 alleles. Rp5 also mapped distally to most Rp1 alleles. Other aspects of recombination at Rp1 suggested that some alleles carry duplicated sequences, that mispairing can occur, and that unequal crossing-over may be a common phenomenon in this region; susceptible progeny from an Rp1 A homozygote had recombinant flanking marker genotypes, and susceptible progeny from an Rp1 DlRp1 F heterozygote showed both possible nonparental flanking marker genotypes.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00260649

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Analysis of the chromosomal distribution of transposon-carrying T-DNAs in tomato using the inverse polymerase chain reaction (1994)

Thomas, Colwyn M. ; Jones, David A. ; English, James J. ; [et al.]

Springer

Molecular genetics and genomics 242 (1994), S. 573-585

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ISSN: 1617-4623

Keywords: Tomato ; Agrobacterium T-DNA ; Inverse PCR ; Activator ; Transposon tagging

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract We are developing a system for isolating tomato genes by transposon mutagenesis. In maize and tobacco, the transposon Activator (Ac) transposes preferentially to genetically linked sites. To identify transposons linked to various target genes, we have determined the RFLP map locations of Ac- and Dissociation (Ds)-carrying T-DNAs in a number of transformants. T-DNA flanking sequences were isolated using the inverse polymerase chain reaction (IPCR) and located on the RFLP map of tomato. The authenticity of IPCR reaction products was tested by several criteria including nested primer amplification, DNA sequence analysis and PCR amplification of the corresponding insertion target sequences. We report the RFLP map locations of 37 transposon-carrying T-DNAs. We also report the map locations of nine transposed Ds elements. T-DNAs were identified on all chromosomes except chromosome 6. Our data revealed no apparent chromosomal preference for T-DNA integration events. Lines carrying transposons at known map locations have been established which should prove a useful resource for isolating tomato genes by transposon mutagenesis.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00285281

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Homologous recombination between plasmid DNA molecules in maize protoplasts (1991)

Lyznik, Leszek A. ; McGee, J. David ; Tung, Po-Yen ; [et al.]

Springer

Molecular genetics and genomics 230 (1991), S. 209-218

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ISSN: 1617-4623

Keywords: Homologous recombination ; Protoplast transformation ; β-Glucuronidase ; Maize

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Summary The requirements for homologous recombination between plasmid DNA molecules have been studied using the PEG (polyethylene glycol)-mediated transformation system of maize (Zea mays L.) protoplasts coupled with the transient expression assay for β-glucuronidase (GUS). Two plasmids were introduced into maize protoplasts; one plasmid (pB×26) contained a genomic clone of the Adh1 maize gene; the other plasmid (piGUS) was a promoterless construction containing part of intron A of the Adhl gene fused to the gusA coding sequence. Thus, the two vectors shared an effective homologous region consisting of a 459 by (Hindlll—PvuII) fragment of the yAdh1 intron A sequence. An active gusA fusion gene would result upon homologous recombination between the plasmids within the intron A sequence, and indeed GUS activity was observed in extracts following co-transformation of maize protoplasts with the two plasmids. The presence of recombinant DNA molecules in protoplast DNA isolated 1 day after co-transformation was verified using polymerase chain reactions (PCR) and Southern blots. For efficient homologous recombination, both plasmids had to be linearized. The recombination reaction was induced by restriction of the plasmid molecules either inside the effective homologous region or at the borders of the intron sequence. However, the presence of even small, terminal, nonhomologous sequences at the 3′ end of the pB×26 fragment inhibited the recombination reaction. Also, both ends of the linearized piGUS DNA molecules were involved in the recombination reaction. The results revealed some features of homologous recombination reactions occurring in plant cells which cannot be accommodated by mechanisms postulated for similar reactions in animal system and in lower eukaryotes.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00290670

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