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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 16 (1989), S. 95-98 
    ISSN: 1432-0983
    Keywords: Kluyveromyces ; Transformation ; 2μ-like plasmids ; G418 resistance ; pKDI
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary pKDI is a 2μ-like circular plasmid found in the yeast Kluyveromyces drosophilarum that can also stably replicate in Kluyveromyces lactis. We have found a short intergenic region in this genome that appears to be functionally neutral; that is, the introduction of foreign sequences into the single EcoRI restriction site located near one of the inverted repeats did not affect the high stability of the natural plasmid. By introducing a G418 resistance gene at this site, we constructed an autonomous recombinant plasmid. Since this vector did not require cir + hosts for its stable maintenance, it could be used to examine the transformation host range of pKD1 among all the species belonging to the genus Kluyveromyces. Both species closely related to K. drosophilarum as well as a few other species that are very different in chromosomal GC % could be transformed to yield highly stable transformant clones.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Kluyveromyces lactis ; Transformation ; pKD1 ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The host specificity of the 2μ-like circular plasmid pKD 1 is such that this plasmid replicates stably in several species of Kluyveromyces yeasts, but not in Saccharomyces cerevisiae. pKD 1-derived plasmids containing various parts of the pKD 1 sequence were capable of transforming Kluyveromyees lactis with high efficiency. When such vectors were introduced into host strains that contained resident pKD1 plasmid, the input DNA frequently recombined with it to produce high proportions of additive recombinant molecules that replicate stably. Recombination events were shown to occur with vectors differing for the presence or absence of the putative origin of replication and of the inverted repeats. Structure, stability and copy number of the recombination products were analyzed for various types of vectors.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0983
    Keywords: Yeast plasmid ; Transformation ; pKD1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The circular plasmid pKD1 (or 1.6 μm DNA) has recently been isolated from Kluyveromyces drosophilarum. This plasmid appears to have a functional organization analogous to that of the 2 μ DNA of Saccharomyces cerevisiae, although the respective nucleotide sequences show little homology. pKD1 can be transferred to Kluyveromyces lactis where it is replicated stably. Using recombinant molecules derived from pKDl, a practical transformation system has been developed for Kluyveromyces lactis, with an efficiency and stability comparable to the 2 μ-based Saccharomyces cerevisiae transformation system.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 7 (1989), S. 639-643 
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It has been reported that long term cell cultures from maize endosperms are not completely de-differentiated, maintaining some tissue-specific synthesis. We analyzed the expression of zein (the major storage protein of maize seeds) in cultures derived from wildtype and opaque-2 maize endosperms. In wildtype cultures, our data indicate a severe restriction in zein accumulation, resulting both from reduction of transcription and from post-transcriptional events. No detectable zein proteins and trace amounts of transcripts were found in opaque-2 cultures, which do not therefore exhibit a distinctive opaque-2 phenotype.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 11 (1988), S. 203-214 
    ISSN: 1573-5028
    Keywords: endosperm development ; DNA methylation ; storage protein genes ; tissue-specific transcription ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We investigated the methylation state of a set of storage protein genes of maize, coding for zeins and glutelins, in different somatic tissues and in developing endosperms. These genes, present as multigene families in the maize genome and organized in clusters on different chromosomes, are coordinately and specifically transcribed only in endosperm cells. Southern blot analysis of DNA digested with methylation-sensitive restriction enzymes shows a specific and extensive undermethylation of zein and glutelin sequences in the endosperm, while a common methylated pattern is detected in the different somatic tissues and in the embryo. However, a constant fraction of endosperm DNA (∼35%) is methylated at all zein sequences, which are found to be heavily modified in pollen DNA as well. Undermethylation is extended along a zein cluster and cannot be explained by reduced levels of 5-methylcytosine in endosperm DNA with respect to other tissues. The undermethylated state of storage protein genes is already established at an early stage of endosperm development, when transcripts levels for both genes are almost undetectable.
    Type of Medium: Electronic Resource
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