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At least 19 recordsLinked to original sources

Genetic analysis of tumorigenesis: IV. Chromosome reduction and marker segregation in progeny clones from Chinese hamster cell hybrids.

Hybrid cells produced by the fusion of pairs of cells, one a tumorigenic derivative of CHEF/16 and the other a nontumorigenic derivative of CHEF/18, give rise to clones which are largely tetraploid, but rare reduced hybrids with chromosome counts in the diploid range have been recovered from tumors of hybrid origin. This paper describes the recovery in cell culture of reduced hybrids in the diploid range by selection with 5-bromodeoxyuridine (BrdU) or methylcellulose as well as by growth in culture of cells from excised tumors. All selected subclones were tumorigenic and resistant to BrdU, but they segregated for resistance to 6-thioguanine. Unselected subclones were tetraploid, nontumorigenic, and sensitive to both drugs. These data show that chromosome reassortment as well as extensive chromosome reduction both occur in a small fraction of the population during growth of each hybrid clone.

Animals

Fate of mismatched base-pair regions in polyoma heteroduplex DNA during infection of mouse cells.

Heteroduplex DNA has been constructed from two variants of polyoma virus that differ genotypically at four distinct sites. The genotypes of the progeny virus derived from infections of mouse cells with single heteroduplexes have been analyzed to determine how the genotypic markers of the parental heteroduplex segregate. Markers that are separated by a length of DNA greater than 600 nucleotides segregate independently. Segregation was not detected between two markers separated by only approximately 90 nucleotides. We interpret these results on the basis of the correction of mismatched base-pair regions in the heteroduplex before the completion of DNA replication. We suggest that this technique provides valuable information concerning gene conversion in mamalian cells and permits the transfer of genotypic markers from one virus strain to another.

Base Sequence

Physiological factors involved in the transformation of Mycobacterium smegmatis.

Transfer of streptomycin resistance and changes from methionine and leucine auxotrophy to prototrophy were achieved in Mycobacterium smegmatis by transformation. Recipient cells were more resistant to mitomycin C and methyl methlanesulfonate treatments than were wild-type cells. A high level of calcium ions was essential for transformation, especially during DNA adsorption, whereas the presence of magnesium ions and the exposure of recipient cells to mild doses of UV light enhanced recombination frequencies. Transformants were not isolated when recipient cell-DNA mixtures were first treated with deoxyribonuclease. Recipient cells at various stages of growth showed similar transformabilities. Transformation was successful only when recipient cells were incubated on rich agar medium after mixture with DNA. Exposure of recipient cells to Pronase before treatment with donor DNA did not affect transformation, suggesting the absence of a protein competence factor. Throughout the present experiments, cotransformation frequencies were very low and unselected-marker segregation patterns were independent, indicating that the methionine, leucine, and streptomycin markers are not closely linked in M. smegmatis.

Calcium

Segregation studies in CHO hybrid cells: I. Spontaneous and mutagen-induced segregation events of two recessive drug-resistant loci.

The process of segreation or phenotypic expression of two recessive drug-resistant loci from heterozygous Chinese hamster ovary hybrid lines is examined. The spontaneous segregation rates of phytohaemagglutinin resistance (Phar) and a temperature-dependent 8-azaguanine-resistant locus (Azarts) from heterozygous quasitetraploid lines using Luria-Delbruck fluctuation analysis were 5 X 10(-5) and 10(-5) events/cell/generation, respectively. In quasihexaploid lines, the latter rates increased 40- and 200-fold, respectively, and were dependent on the number of presumptive drug-sensitive allelel. The mutagens EMS, MNNG, ICR-170, ICR-191, and gamma rays significantly increased the frequency of segregation events. The mutagen-induced frequency of dominant mutations to ouabain (Ouar) and alpha-amanitin (Amar) rsistance in the same hybrid line was much lower in comparison to segregation events and was mutagen specific. The chromosome number per metaphase cell was more variable than DNA content in quasitetraploid lines. These properties of marker segregation are consistent with mechanisms of either restricted chromosome loss, rearrangement, or mutation.

Azaguanine

Assignment of a gene for human quinoid-dihydropteridine reductase (QDPR, EC 1.6.5.1) to chromosome 4.

An acrylamide gel electrophoretic procedure is described which allows the separation of human quinoid-dihydropteridine reductase (QDPR), EC 1.6.5.1) from the homologous enzyme expressed in established rodent cell lines. The human enzyme marker segregates exclusively with chromosome 4 in a series of well characterized man-mouse somatic cell hybrid clones from our clone bank. This observation supports the assignment of a structural gene for QDPR to human chromosome 4.

Animals

Genetic recombination in fused spheroplasts of Providence alcalifaciens.

Spheroplasts of Providence alcalifaciens strain P29 auxotrophs were prepared by combined treatment with glycine and lysozyme-EDTA. About 15% of spheroplasts had areas of cytoplasmic membrane exposed where cell wall was absent. The spheroplasts of different auxotrophs were mixed pairwise and fusion was attempted with polyethylene glycol or nascent calcium phosphate. After spheroplasts had regenerated to bacterial forms selection was made for recombinants. Recombinants arose at frequencies of 3.8 X 10(-6) to 1.7 X 10(-7) per spheroplast initially present, by both methods of fusion. The frequency was strongly dependent on the number of chromosomal loci used in selection. The possible order of five loci was determined and this corresponded to that on the closely related Proteus mirabilis chromosome. Control experiments excluded possibilities of auxotrophic reversion, conjugation, transformation, transfection or transduction as explanations of the results. Analysis of prototrophic clones yielded stable prototrophs or mixtures of stable prototrophs and stable recombinants. Parental types were not encountered. Unselected markers segregated among recombinants. It was concluded that the formation of recombinant bacteria was due to spheroplast fusion and that only stable products of the very temporary heteroploid state were haploid recombinants. The low frequency of recombination was ascribed to the limited number of spheroplasts with areas of exposed cytoplasmic membrane.

Cell Wall

Idiotypic analysis of the response of C57BL/6 mice to the (4-hydroxy-3-nitrophenyl)acetyl group.

Strain C57BL/6 mice produce a highly restricted primary response to the (4-hydroxy-3-nitrophenyl)acetyl (NP) group. This response is composed of molecules having mu or gamma1 heavy chains and light chains of the lambda type. A guinea pig anti-idiotype was raised to the purified C57BL/6 primary anti-NP immunoglobulin. After suitable absorption, this anti-idiotype was shown to detect markers present on the primary anti-NP immunoglobulin only of those strains which express the Ig-1b allotype. Breeding experiments demonstrated that the marker segregated with the heavy chain linkage group.

Adjuvants, Immunologic

Genetic and functional characterization of an antiserum to the lipid A-specific triggering receptor on murine B lymphocytes.

The inheritance of responsiveness to lipopolysaccharide (LPS), and of a marker recognized in LPS-reactive cells by a heterologous antiserum, was studied in crosses between C3H/HeJ (nonresponder) and C3H/Tif (high responder) mice. F1 hybrid mice show codominant expression of these traits: (a) LPS-reactive cells are only hlaf as frequent in the hybrids as in the high responder parent; (b) the serologically defined marker is expressed in half as many cells in the hybrids as in the high responder parent. In backcross generations, both LPS responsiveness and this serological marker segregated into high, intermediate, and nonresponders. LPS or free lipid A, but not two other B cell mitogens (lipoprotein, and purified protein derivative of tuberculin), compete with the antiserum for binding to the B cell surface membrane, and are capable of completely inhibiting such binding without interfering with the binding of a-ti-Ig antibodies or complexes to Fc receptors. The addition of an IgG fraction of the antiserum to B cell cultures results in exponetial growth of the cells and in maturation to antibody secretion. This mitogenic activity is dose-depedent and absorbable on spleen cells from LPS high responder mice. Taken together, these observations suggest that this antiserum contains antibodies to the lipid A-specific triggering receptor on B lymphocytes.

Animals

Detection of aberrant nuclear DNA metabolism in a conditional mutant of Saccharomyces cerevisiae.

A single recessive nuclear gene mutation has been isolated from strain 123.1C of Saccharomyces cerevisiae which appears to be conditionally deficient in nuclear DNA metabolism. Growth of the mutant strain at the elevated temperature of 36 degree C results in rapid loss of cell viability. However, no apparent reduction in the rate of radioisotope incorporation into DNA was detected during this period. When haploid cells carrying this temperature sensitive lesion were exposed to the restrictive temperature for varying lengths of time, returned to the permissive temperature, mated with a non-temperature sensitive strain and then the resulting diploids made to undergo meiosis, a greatly reduced number of viable spores were produced. Genetic analysis of the viable spores produced by these diploids has revealed aberrant auxotrophic marker segregation patterns. Thus, these results suggest that the mutated gene hardbored in this strain plays a vital role in the metabolism of the nuclear genome.

DNA

Independent segregation of two functional markers expressed on the same B-cell subset in the mouse: the Mls determinants and LPS receptors.

Mice of the C3H/Tif strain display a mixed leukocyte reaction (MLR) with all H-2k strains carrying any of the known alleles of the Mls locus. In particular, C3H/Tif is incompatible with the related substrain C3H/HeJ, from which it also differs at the locus responsible for the recognition of lipopolysaccharides (LPS) as B-cell mitogens, and at the Mod-1 locus. Our genetic analysis indicates that the MLR incompatibility between these strains is not H-2-linked and segregates as controlled by a single locus, most probably identical to Mls, for which the C3H/Tif strain expresses a previously unidentified allele, Mlse. Moreover, segregation data show that this locus assorts independently of LPS responsiveness and that neither marker is closely linked to the Mod-1 locus in linkage group II.

Alleles

Plasmid replication and Hfr formation in strains of Escherichia coli carrying seg mutations.

Several conditional-lethal mutantions that do not permit the replication of F-factors of Escherichia coli K-12 are located at a site called seg. This gene is located on the E. coli chromosome between ser B and thr. It is unrelated to other known genes involved in DNA replication. Strains carrying seg mutations were unable to replicate F'-lac+, several F'-gal+s, F'-his+ and bacteriophage gamma at 42 degrees. However, neither phage T4, ColE1, nor any of the R factors tested were prevented from replicating at 42 degrees C. When the kinetics of the loss of F-primes is studied in seg strains, it is found that the rate of curing depends on the size of the plasmid, larger F factors curing faster than smaller ones, and that Hfrs are formed at high frequencies. The Hfrs showed both F-genote enlargement and normal transfer of chromosomal markers. The F-genotes are unstable and segregate chromosomal markers at high frequencies. Some orthodox Hfrs were examined, and two that were known to revert to the F+ condition relatively frequently were found to generate enlarged F-genotes on mating, whereas two strains that were very stable with respect to the F+ state did not show F-genote formation. F-genote formation from seg Hfr stains is dependent on a functional recA gene, as F-genote formation was not seen with a seg-2, recA-1 Hfr. This is in contrast to F-genote enlargement shown by both orthodox Hfrs and an Hfr strain constructed by integration of a temperature-sensitive F'-gal+, whose F-genote enlargement is Rec-independent. Thus there may be more than one mechanism for the formation of enlarged F-genotes.

Chromosome Mapping

High frequency of mutation to tubercidin resistance in CHO cells.

The acquisition of high-level resistance to tubercidin (an adenosine analog) in CHO cells occurs in a single step at high frequency (10(-3) to 10(-4)) without mutagenesis. Analysis of a large number of independent mutants by a fluctuation test (Luria and Delbruk, 1943) indicates that they arise independently of the selection medium and all fall into the same complementation group. All mutants tested lack detectable adenosine kinase activity. An analysis of hybrids between mutant and wild-type cells indicates that resistance to tubercidin is a recessive marker which segregates as would be expected if it were a haploid locus in the parental CHO cell. Resistance to tubercidin is not linked to the X chromosome in CHO cells and appears to occur at much lower frequency in primary Chinese hamster cells and other cultured cell lines.

Animals

Locating salmonella resistance gene on mouse chromosome 1.

The inherited resistance of inbred mouse strains to Salmonella typhimurium injected subcutaneously has been reported to be controlled primarily by a single gene designated Ity. Resistant mice have the dominant allele Ityr and sensitive mice are homozygous for the recessive allele Itys. This paper describes studies undertaken to locate the gene using readily distinguishable phenotypes as chromosome markers. Appropriate F1 backcross and F2 generations of hybrids between resistant and susceptible inbred strains of mice, with or without the particular phenotypic markers, were tested both for susceptibility to salmonella sci and presence of the marker. Independent segregation of the characteristics eliminated all chromosomes except Chromosome 1. C57L mice resistant to S. typhimurium, Ityr Ityr and leaden, ln ln, located on Chromosome 1, were crossed with BALB/c mice sensitive to S. typhimurium and non-leaden. In the F2 generation mice, ln always segregated with Ityr. The presence of only one leaden mouse sensitive to S. typhimurium out of sixty leaden F2 mice tested linked Ityr closely with ln on Chromosome 1. This result will be of use in further experiments with hybrid populations by enabling us to predetermine resistance or sensitivity to S. typhimurium without infecting the mice, so permitting experiments on the nature of the inheritance in unsensitized mice.

Animals

Genome-wide association identifies and validates genomic region controlling grain yield and agronomic traits in extra-early orange maize inbred lines under drought.

In order to meet the expected maize yield by 2050, breeders must work to improve breeding program efficiency by intensifying the implementation of new and improved technologies such as marker-assisted selection (MAS). Dissecting the genomic regions associated with drought tolerance is the first step forward in MAS program deployment for maize improvement under drought stress. Genome-wide association studies (GWAS) were used to investigate and identify quantitative trait loci (QTLs) associated with six traits under drought stress. One hundred and eighty-seven extra-early orange maize inbred lines were evaluated under managed drought stress at Ikenne, in Nigeria, during the 2022 and 2023 dry seasons. The materials were also genotyped using 9355 DArTseq SNP markers and analyzed using the enriched compressed mixed linear model (ECMLM). Enriched compressed mixed linear model was used for association-trait analysis. The ECMLM-based GWAS identified 45 candidate genomic loci associated with the six traits, including five for grain yield, with R2 ranging from 8.79 to 25.3%. Independent validation using the multi-locus 3VmrMLM approach confirmed seven high-confidence genomic loci consistently detected by both methods across grain yield, anthesis-silking interval, ear aspect, and ears per plant, providing additional statistical support for these genomic regions. Candidate gene annotation identified biologically relevant genes underlying the validated loci, including Zm00001eb238250 (protein-serine/threonine phosphatase), Zm00001eb040940 (trehalose-phosphatase), Zm00001eb117820 (homeobox protein knotted-1-like 4), Zm00001eb145560 (zinc ion-binding protein), and Zm00001eb294180 (WRKY DNA-binding domain protein), suggesting their potential roles in drought adaptation and grain productivity. These findings improve our understanding of the genetic architecture of drought tolerance in extra-early orange maize and provide valuable genomic resources for accelerating drought-resilient maize breeding.

Zea mays

Transfer of multiple antibiotic resistance from Bacteroides fragilis to Escherichia coli.

Multiple antibiotic resistance was transferred from a clinical isolate of Bacteroides fragilis to a strain of Escherichia coli K12. Resistance to ampicillin, amoxicillin, cephalothin, tetracycline, minocycline, and chloramphenicol was transferred as a unit, but the resistance markers became segregated during storage of the recipient strains.

Bacteroides fragilis

Segregation of an internal biochemical marker during influenza virus recombination and its possible correlation with biological properties.

The internal matrix proteins of A/Okuda/57, A/Finland/4/74 and A/New Jersey/8/76 viruses and several recombinant strains have been examined by radioiodination of the purified proteins followed by peptide mapping. The method is rapid and requires only small amounts of material. Reproducible differences were detected between the matrix proteins of the above parents and allowed the origin of the matrix proteins of the recombinant viruses to be determined. The use of matrix protein identity as a marker in recombination work and its possible correlation with biological properties of the virus is discussed.

Crosses, Genetic

Linkage of tobiano coat spotting and albumin markers in a pony family.

Genetic segregation patterns among blood type markers and various phenotypically observed traits were studied in a small herd of ponies. The herd consisted of 10 mares without white spotting and a single stallion with the dominant pattern of tobiano spotting. Comparison of segregation patterns at loci for which the stallion was heterozygous showed tight linkage for the Alb-B and tobiano markers. In 17 cases in which the Alb contribution of the sire could be determined, all 10 foals that inherited AlbB from him were tobiano spotted, and all 7 non-spotted foals inherited his AlbA. The use of the symbol To is proposed for dominantly inherited tobiano spotting linked to the albumin.

Animals