Unbiased transmission of mutant alleles at the human retinoblastoma locus

The preferential transmission of the mutant allele to offspring from fathers who carry a germline mutation in the retinoblastoma gene was examined by analyzing 46 consecutive pedigrees. Among 75 offspring from 29 fathers, the ratio of carriers to noncarriers was 49%. Among the 106 offspring from 55 mothers the ratio was 57%. Neither ratio differs statistically from the expected 50%. When the analysis was limited to only those families with low-penetrance retinoblastoma, we still did not observe a biased transmission of alleles from fathers, although mothers did have an excess of carrier offspring of borderline statistical significance (the P-value was approximately 0.03). While we cannot rule out a biased transmission of alleles from some parents, there appears to be no such bias overall.     

Structure of mutant alleles at the aprt locus of Chinese hamster ovary cells

To determine the types of gene structural alterations causing deficiency of adenine phosphoribosyl transferase (aprt) activity in spontaneous and chemically induced mutations of cultured somatic cells, we analyzed the restriction enzyme cleavage patterns of aprt gene sequences in mutant strains selected from Chinese hamster ovary cells. Patterns of aprt-containing fragments in Southern blots were mostly unchanged in our collection of 280 ethyl methane sulfonate-induced and spontaneous aprt- mutants, suggesting that base-pair changes or other alterations below our limit of resolution on agarose gels (approximately 50 base-pairs) are responsible for the great majority of mutations at the aprt locus. Occasionally, these mutations could be localized when they resulted in the loss or gain of a restriction enzyme site and the generation of new fragments of predictable size. Deletions of aprt-containing sequences were detected in only eight of 119 spontaneous mutants and in only one ethyl methane sulfonate-induced mutant. An insertion of 300 base-pairs near the 5' end of the aprt structural gene was found in one spontaneous aprt- strain. This insertion mutant was stable with a reversion frequency of less than 2 X 10(-7). Several unstable aprt- mutants were detected in our collection, but these had no observable alterations of aprt coding or flanking sequences.     

Regulation of white locus expression: the structure of mutant alleles at the white locus of Drosophila melanogaster

We have analyzed the structures of 19 mutant alleles at the white locus of Drosophila melanogaster. Thirteen of the mutant alleles in our selected sample arose spontaneously, and of these, seven are associated with insertions of non-white-region DNA sequence elements. Several lines of evidence strongly suggest that these insertions are responsible for their associated mutant alleles, and further suggest that most or all of these insertions are transposons. Moreover, the white locus DNA sequences can be divided into two nonoverlapping domains on the basis of the properties of the two domains as mutational targets. One of these domains behaves, in this regard, in the manner expected of functional coding sequences, whereas the other does not. We propose a model for the nature and function of the presumptive noncoding white locus genetic elements. The two domains of the white locus defined by our studies are approximately coextensive with the functionally distinct subintervals of the locus defined by previous genetic analysis. Lastly, our results strongly suggest that the dominant, mutable wDZL allele results from the insertion of a transposon outside of, but near, the white locus. This putative transposon apparently carries genetic elements that act at a distance to repress expression of the white locus.     

Four new alleles at the mannose-6-phosphate isomerase locus in rabbit

Four new alleles at the rabbit ( Oryctolagus cuniculus ) mannose-6-phosphate isomerase (E. C. 5. 3. 1. 8, MPI) locus are proposed to account for phenotypes observed after starch gel electrophoresis and enzymatic staining of red cell lysates and tissues. Population data from various wild and domestic rabbit populations are presented.     

Characterization of two new alleles at the goat CSN1S2 locus

Two novel alleles at the goat CSN1S2 locus have been identified: CSN1S2(F) and CSN1S2(D). Sequence analyses revealed that the CSN1S2(F) allele is characterized by a G --> A transition at the 13th nucleotide in exon 3 changing the seventh amino acid of the mature protein from Val to Ile. The CSN1S2(D) allele, apparently associated with a decreased synthesis of alpha s2-casein, is characterized by a 106-bp deletion, involving the last 11 bp of the exon 11 and the first 95 bp of the following intron. Methods (PCR-RFLP and PCR) for identification of carriers of these alleles have been developed.     

The generation of new S alleles at the incompatibility locus of Lycopersicum peruvianum Mill

Summary A detailed analysis has been made of S genotypes in progenies derived from induced and spontaneous inbreeding processes in a clonal population of Lycopersicum peruvianum Mill. The results indicate that, in certain genetic backgrounds, induced inbreeding leads to the generation of a new S allele which usually first appears in the pistil of individuals otherwise homozygous for one of the parental specificities. When the change in specificity occurs in S heterozygotes, spontaneous self-compatibility is promoted and the new allele can be transmitted, via selfing, to the following generation.The factors and mechanisms which may be involved in the generation of new specificities at the S locus of higher plants are discussed and preliminary evidence is provided which suggests that the hypothesis of mutation by equal crossing-over is not applicable to the present study.

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Zusammenfassung Es wurde eine ausführliche Analyse der S-Genotypen in Nachkommenschaften einer geklonten Population von Lycopersicon peruvianum Mill, nach induzierter und spontaner Inzucht gemacht.Die Ergebnisse deuten an, da induzierte Inzucht bei einem bestimmten genotypischen Milieu zum Entstehen eines neuen S-Allels führt. Dieses erscheint gewöhnlich zuerst in den Griffeln einzelner Individuen, die im übrigen für eines der elterlichen Allele homozygot sind. Wenn die Änderung in S-Heterozygoten auftritt, dann wird die spontane Selbstkompatibilität gefördert. Das neue Allel kann durch Selbstung in die nächste Generation übertragen werden.Die Faktoren und Mechanismen, die am Entstehen neuer Allele am S-Locus höherer Pflanzen beteiligt Scin können, werden diskutiert. Ein vorläufiger Hinweis wird dafür erbracht, da die Hypothese der Mutation durch equal crossing-over für die vorliegende Untersuchung nicht zutrifft.

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This publication is contribution no. 644 of the EURATOM Biology Division.     

Generation of new Notch2 mutant alleles

The Notch signaling pathway is an evolutionarily conserved intercellular signaling mechanism, and mutations in its components disrupt embryonic development in many organisms and cause inherited diseases in humans. We previously described construction and analysis of a hypomorphic allele of the Notch2 gene. Homozygosity for this allele leads to embryonic and perinatal lethality due to cardiovascular and kidney defects. We report here novel Notch2 mutant alleles generated by gene targeting in embryonic stem cells, including a conditional null allele in which exon 3 of the Notch2 gene is flanked by loxP sequences. These new Notch2 mutant alleles expand the set of tools available for studying the myriad roles of the Notch pathway during mammalian development and will enable analysis of Notch2 function at additional stages of embryogenesis and in adult mice.     

Fertile heteroallelic combinations of mutant alleles of the otu locus of Drosophila melanogaster

Summary This paper describes the ovarian pathologies observed when 108 different heteroallelic combinations were made involving 17 independent mutations at the ovarian tumor (otu) locus. Most of the mutant phenotypes can be explained as graded responses by individual germ cells to different levels of functionally active otu gene product (OGP) synthesized by the mutant cells themselves. The lowest and highest levels of OGP appear to be produced by otu ¹⁰ and otu ¹⁴, respectively. In most heteroallelic ovaries the alleles have additive effects, and hybrid germ cells reach a developmental stage more advanced than the weaker homozygote but less advanced than the stronger homozygote. However, examples of both positive and negative complementation also have been found, and these suggest that the products encoded by different mutant alleles can combine to form dimers or multimers which may be superior or inferior to the homodimers. In flies homozygous for otu ¹¹ most ovarioles contain tumors, but some germ cells are able to develop further than those in otu ¹⁴ homozygotes. This suggests that, while otu ¹¹ produces intermediate levels of OGP, it also produces a second product (which otu ¹⁴ cannot make) that is utilized at the period in oogenesis when development in cells homozygous for otu ¹⁴ is blocked. When otu ¹¹ is combined with any one of eight specific alleles, it allows oocyte/nurse cell syncytia to differentiate that can complete development and undergo embryogenesis, if fertilized. The endopolyploid nurse cells of these hybrids have giant polytene chromosomes, and the presence of GPCs in functionally active, germ-line derived cells provides an interesting new system for experimental study. Analysis of the characteristic ovarian pathologies produced by flies of different genotypes leads to the conclusion that the products of the otu ⁺ gene are utilized during at least six different periods in Drosophila oogenesis.     

A new mutant at the Dystrophia muscularis (dy) locus in the SM strain of mice

Muscular dystrophy was found in the SM strain of mice. This defect was shown to be caused by a single autosomal recessive gene allelic with the genes at the dy locus. The dystrophic SM mice may provide a useful animal model for human muscular dystrophy, because SM strain has been selected for small body size and carries rare alleles at several loci.     

Chromosome landing at the tomato Bs4 locus

The tomato (Lycopersicon esculentum) Bs4 gene confers resistance to strains of Xanthomonas campestris pathovar vesicatoria that express the avirulence protein AvrBs4. As part of a map-based cloning strategy for the isolation of Bs4, we converted Bs4-linked amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) markers into locus-specific sequence-tagged-site (STS) markers. The use of these markers for the analysis of 1972 meiotic events allowed high-resolution genetic mapping within a 1.2-cM interval containing the target gene. Two tomato yeast artificial chromosome (YAC) clones, each harboring inserts of approximately 250 kb, were identified using the marker most closely linked to Bs4. YAC end-specific markers were established and employed to construct a local YAC contig. The ratio of physical to genetic distance at Bs4 was calculated to be 280 kb/cM, revealing that recombination rates in this region are about three times higher than the genome-wide average. Mapping of YAC end-derived markers demonstrated that the Bs4 locus maps within a region of 250 kb, corresponding to a genetic interval of 0.9 cM.     

"Silent" alleles at the HLA-C locus.

Defined specificities of the HLA-C locus possess a significantly lower immunogenicity as compared with the HLA-A and HLA-B specificities. HLA-C "blanks" are not immunogenic at all. Among the possible explanations, the most likely one appears to be a low phenotypic expression of these alleles. C locus antigens as a whole are thus likely to be of minor relevance in donor/recipient matching for transplantation.     

Superunstable alleles at the cut locus in Drosophila melanogaster

Summary This is a detailed study of the reversions of the ct ^MR2 allele putatively carrying á mobile element (MR-transposon) in the cut locus. Stable, unstable and superunstable revertants have been identified. Besides, a series of multiple unstable visible and lethal ct mutations derived from the ct ^MR2 allele have been obtained. They are shown to include supermutable alleles. The results suggest that the MR-transposon is connected with at least three functions: excision; change of orientation; and change of position within the cut locus, these functions being disturbed in different ways in different unstable ct ⁺ and ct alleles. In some cases the mutant transitions are somehow strongly stimulated leading to superinstability, reaching the rate of 0.5.     

Integration efficiencies of spontaneous mutant alleles of amiA locus in pneumococcal transformation

The distribution of integration efficiencies of independent mutations spontaneously occurring in the amiA locus was determined in two strains of pneumococcus. Strain Cl(3) integrates genetic markers with different efficiencies during transformation, whereas strain 401, isogenic with strain Cl(3), does not discriminate between markers and integrates all markers with the same high efficiency. The discriminating strain Cl(3) gives rise to spontaneous mutations in the locus amiA, which fall into four classes with respect to their individual integration efficiencies. Mutations with a low efficiency of integration are equal in number to mutations with a high efficiency. Mutations from the two other classes corresponding to intermediate and very high efficiencies are about five times less frequent. The same four classes were also found among amiA mutants spontaneously occurring in strain 401. However, the two distributions of integration efficiencies of amiA mutants arisen either in strain Cl(3) or strain 401 are significantly different. The number of spontaneous amiA mutants, estimated by two methods, was found to be higher in strain 401 than in strain Cl(3). The increase of the mutation rate in strain 401 could be accounted for by the excess of mutations falling in the two less-efficient classes observed in this strain. The discriminating process which acts during transformation presumably also intervenes in the appearance of spontaneous mutations.