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Biomedical subjects

H Saedler

Publications and source records attributed to H Saedler.

At least 109 records · Page 6Linked to original sources
PubMed1986

Molecular analysis of the En/Spm transposable element system of Zea mays.

The nucleotide sequence of the autonomous transposable element En-1 isolated from the wx-844::En-1 allele has been determined. En-1 is 8287 bp long. The structure of the mosaic gene 1, coding for the major En transcript, has been established. The promoter gene 1 is located in the highly structured left end of the element and the gene spans almost the entire length of En-1. The first intron of gene 1 is 4434 nucleotides long and contains two large open reading frames, 2714 bp and 761 bp in size, which hybridize to minor RNA species in Northern blot experiments.

Alleles↗
PubMed1985

Genetic and molecular analysis of the Enhancer (En) transposable element system of Zea mays.

A newly isolated, unstable mutation wx-844::En-1 of Zea mays was proven to be caused by the insertion of the autonomous transposable element En into the Waxy (Wx) gene. Molecular analysis revealed that En-1 is 8.4 kb long, has a 13-bp long perfect inverted repeat at its termini and generates a 3-bp target site duplication. En-1 is integrated into an intron located approximately in the middle of the transcribed region of the Wx gene. Structural evidence is presented indicating that a receptor component (Inhibitor) can arise by internal deletion of an autonomous En element.

Alleles↗
PubMed1985

Molecular interactions between the components of the En-I transposable element system of Zea mays.

The sequence of the Inhibitor element Spm-I8 isolated from the wx-m8 allele has been determined. The element is 2242 bp in length. Its ends can be folded into long stem and loop structures. In a line containing the autonomous En element (wx-m8+En) we have detected a 2.5-kb transcript hybridizing to Spm-I8. A cDNA copy of this En-specific transcript containing 1.2 kb of the 3' end was cloned and it DNA sequence was determined. The 3' half of the cDNA is homologous to Spm-I8 and the region of homology is interrupted by intervening sequences. In the absence of an autonomous En element two chimeric transcripts are observed in the wx-m8 line which are probably initiated at the wx promoter and terminate in the Spm-I8 insertion. In the presence of the En element, these transcripts are suppressed, possibly by a trans-acting function of En, inhibiting transcription read-through into Spm-I8.

Alleles↗
PubMed1985

Transposition in plants: a molecular model.

A molecular model for transposition of plant transposable elements is described. This process may occur via excision and re-integration of the element. Excision generates DNA sequence diversity which suggests the participation of DNA repair enzymes in the healing of the donor molecule.

DNA Transposable Elements↗
PubMed1985

Plant transposable elements generate the DNA sequence diversity needed in evolution.

Two germinal and 16 somatic reversion events induced by the Enhancer (En) transposable element system at the wx-8::Spm-I8 allele of Zea mays were cloned and studied by sequence analysis. Excision of the Spm-I8 receptor element from the wx gene results in various mutant DNA sequences. This leads to altered gene products, some of which are still capable of restoring the wild-type phenotype. Possible 'foot-print' sequences that may have arisen by the excision of transposable elements were observed when intron sequences of the wild-type (wx+) and mutant (wx-m8) alleles of the wx gene were compared. The sequence divergence generated by visitation of a locus by plant transposable elements is discussed with respect to the molecular evolution of the new gene functions.

Base Sequence↗
PubMed1985

Molecular cloning of the a1 locus of Zea mays using the transposable elements En and Mu1.

The a1 locus of Zea mays has been cloned using transposable elements as gene tags. The strategy was to make genomic libraries from maize stocks with a1 mutations induced either by En(Spm) or by Robertson's Mutator-system. These libraries were then screened with either Spm-I8 and En1, for the En-containing mutant, or with Mu1 for the Mu-induced mutation. There are many En and Mu1 hybridizing sequences present in the maize genome, however, by a process of cross-screening of the positives from the two libraries and by molecular analysis of the En-positive clones it was possible to identify clones in both libraries carrying all or part of the a1 gene.

Alcohol Oxidoreductases↗
PubMed1985

Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging.

The pal locus of Antirrhinum majus was cloned using the transposable element, Tam 3, as a probe. The pal clone was used to examine, at the molecular level, those aspects of instability previously observed phenotypically and genetically. The effects of temperature and of genetic background on excision of the element at pal are considered, and related quantitatively to the phenotype. We describe the identification of the transcript of the pal locus and show that insertion of Tam 3 blocks the production of a normal pal transcript in developing flower buds.

Journal Article↗
PubMed1985

Sequence comparison of 'states' of a1-m1 suggests a model of Spm (En) action.

Two states of the a1-m1 allele featuring different phenotypes in the absence as well as in the presence of Spm or En have been cloned and sequenced.. The insertion site and orientation of the Inhibitor (I) element within the two alleles is identical. The sizes of the I elements differ, being 2.2 kb in state 6078 and 789 bp in state 5719A-1. The internal deletion in state 5719A-1 affects sequences within one side of the terminal inverted repeats of the I element. This alteration can be correlated with the decreased response of this state to the Mutator function of Spm. A model for the interaction between Spm (En)-encoded functions and the receptor element is discussed explaining the phenotypic differences between the states of the locus.

Alleles↗
PubMed1984

Repetitive sequences and their organization on genomic clones of Zea mays.

Fourteen recombinant clones from Zea mays were studied with regard to their composition of unique and repetitive sequences. Southern hybridization experiments were used to classify restriction fragments of the clones into a unique, middle or highly repetitive class of reiteration frequency. All three classes were often found on the same genomic clone. Crosshybridization studies between clones showed that a given repeat might be present on several clones, and thus four families of highly repetitive elements were established. Heteroduplex analysis was used to show the arrangement and size of repeats common between several clones. A short interspersion pattern of unique, middle and highly repetitive DNA was found. The dispersed repetitive elements were 300-1300 bp in length. Analysis of the pattern produced by a given repeat in genomic Southern experiments suggests that some small dispersed repeats may also exist as part of a larger repeating unit elsewhere in the genome.

Journal Article↗
PubMed1984

The 17-kb Tam1 element of Antirrhinum majus induces a 3-bp duplication upon integration into the chalcone synthase gene.

The DNA sequence of the termini and the flanking regions of the 17-kb transposable element Tam1 was determined. Tam1 is integrated in the chalcone synthase gene of the niv-53 mutant of Antirrhinum majus. The element has a 13-bp perfect inverted repeat at its termini and appears to induce a 3-bp duplication of the target site upon integration. The DNA sequence of a niv revertant was analyzed and found to differ from the wild-type sequence by an additional 2 bp that seem to derive from the target site duplication. Stretches of homologous sequences have been found between the ends of Tam1, within each terminus of the element, and between the termini and target site sequences. Structural similarities between the ends of Tam1 and the Spm-18 element of Zea mays reflect a possible horizontal spread of a common progenitor.

Journal Article↗
PubMed1984

The Spm (En) transposable element controls the excision of a 2-kb DNA insert at the wx allele of Zea mays.

The waxy (Wx) locus of Zea mays was cloned from strains carrying the wild-type and wx mutant alleles. The receptor component of the Suppressor-Mutator (Spm) controlling element system in the wx allele was shown to be a 2 kb long insertion within the transcribed region of the Wx gene. The insertion, termed Spm-I8, is excised during somatic reversion events induced by the autonomous controlling element Enhancer (En), which is an equivalent to Spm. Integration of Spm-I8 into the Wx gene generates a 3-bp target site duplication. Spm-I8 has a 13 bp long inverted repeat at its termini. The ends of the element can be further folded to build a large double-stranded structure consisting of five perfectly matching double-stranded regions of 9-13 bp in length, interrupted by single-stranded loops. A comparison of the wild-type and wx alleles revealed two additional insertions 6 (insert-1) and 0.25 (insert-2) kb in length. No En-induced excision of insert-1 and insert-2 could be detected so far. There is remarkable structure and sequence homology between Spm-I8 and the transposable elements Tam1 and Tam2 of Antirrhinum majus at their termini, reflecting a possible evolutionary and/or functional relationship between transposons in different plant species.

Journal Article↗
PubMed1981

Transposon Tn951 (TnLac) is defective and related to Tn3.

Tn951 is flanked by two perfect inverted repeats of 41 bp which include the 38 bp sequence of the IR of Tn3. Tn951 also contains the last 100 bp of the tnpA gene but with at least two mutations. However, beyond nucleotide 137 the sequences diverge and hybridization experiments show that Tn951 lacks at least the first two thirds of the tnpA gene. In agreement with these observations Tn951 does not transpose by itself at a detectable frequency but can be complemented by the tnpA gene of Tn801 or Tn3. Tn501, Tn1721 and gamma delta do not complement Tn951 transposition. Transposition of Tn951 duplicates 5 bp of target DNA sequence.

Base Sequence↗
PubMed1981

A new type of IS1-mediated deletion.

Genetical tests and DNA sequence analysis revealed that the mechanism of formation of IS1-induced type I and type II deletions differs. IS1-mediated type II deletions occur at the termini of the integrated element and do not remove the element. This process is independent of the cellular recA system and does not involve DNA sequence homology. Conversely, the formation of IS1-induced type I deletions differs substantially. They require recA gene product, small DNA sequence duplications and a topological arrangement of the DNA molecule to allow alignment of duplications.

Base Sequence↗
PubMed1980

Deletions and an inversion induced by a resident IS1 of the lactose transposon Tn951.

DNA-DNA filter binding tests, "Southern" blotting experiments and DNA heteroduplex analysis clearly show that Tn951 contains an IS1 element. This IS1-951 sequence is peculiar in that it does not contain the PstI cleavage site which is usually observed on E. coli derived IS1 elements. Nonetheless, IS1-951 induces deletions. This process is temperature dependent. One instance of an IS1-951 induced inversion was observed, the structure of which is compatible with the current models of transposition of IS elements.

Base Sequence↗