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PubMed1977

Transcription of ribosomal protein genes carried on F' plasmids of Escherichia coli.

The transcriptional activity of the DNA sequences coding for certain ribosomal proteins has been measured in Escherichia coli. Two partial diploid strains were isolated from mating of a recA, argD, aroE recipient and an Hfr with an origin at 69.5 min. One contained an F' element carrying the genes from 69.5 to 72.7 min including the rpsL locus (72.4) and the second was diploid for the genes from 69.5 to 71.5 min but did not include any mapped ribosomal protein loci. The molecular weights of the plasmids were estimated to be 140 and 110 x 10(6) daltons, respectively. The extent of in vivo transcription of the chromosomal genes on the plasmid and the ribosomal protein mRNA fraction of the total cellular RNA weer calculated from DNA-RNA liquid hybridization experiments using both DNA and RNA excess procedures. The results indicated a high degree of transcriptional activity concentrated in the ribosomal protein sequences with 83% of the F' chromosomal sequences transcribed into mRNA products representing about 0.12% of the total cellular RNA.

Conjugation, Genetic↗
PubMed1977

A suitable method for construction and cloning hybrid plasmids containing EcoRI-fragments of E. coli genome.

A convenient procedure for the isolation of specific EcoRI-fragments of E. coli genome and their amplification on Km-resistance plasmid victor CK delta11 is described. The hybrid molecules were constructued in vitro using EcoRI-digestion, followed by ligation. Then appropriated E. coli strain was transformed with ligated DNA mixture and hybrid plasmids CK delta11-arg+, CK delta11-his+, CK delta11-thr+ and CK delta11-leu+ containing loci of E. coli genome were selected by molecular cloning. The hybrid plasmids obtained consisted of one EcoRI-fragment of initial plasmid CK delta11 and one respective specific portion of E. coli genome.

Arginine↗
PubMed1976

Isolation and characterization of Co1E2 plasmid mutants unable to kill colicin-sensitive cells.

After transfer from a mutagenized host, twenty one Co1E2 plasmid mutants were isolated after screening 10,000 clones for abnormal colicin production. Analysis by SDS polyacrylamide slab gel electrophoresis of proteins synthesized after mitomycin C-induction of mutant cultures, indicates that all but two of the mutations are in the structural gene from colicin E2. Of these, nine produce fragments of colicin in both whole cells and minicells and some are suppressed by nonsense suppressors. Studies with a nonsense mutant producing only a small colicin E2 fragment (Co1E2-421) suggest that colicin E2 in not involved in plasmid DNA replication, in the control of its own synthesis, or required for cell death when cells become committed to colicin production. The two plasmid mutants outside the colicin gene segregate plasmid-free cells at 33 degrees, 37 degrees and 43 degrees. One segregates fairly rapidly (about 4% per generation) though the colicin-producing cells make normal amounts of colicin, whilst the other segregates more slowly and the colicin-producing cells make much reduced amounts of colicin.

Colicins↗
PubMed1976

The molecular events involved in the induction of petite yeast mutants by fluorinated pyrimidines.

The fluorinated pyrimidines 5-fluorouracil (5FU) and 5-fluorocytosine (5FC) induce the cytoplasmic petite mutation in the yeast Saccharomyces cerevisiae with high efficiency. It was found that in order to induce the mutation, 5FC must first be deaminated to 5FU. However, mutagenesis does not depend on the further conversion of 5FU to its deoxyriboside (5FUDR) and subsequent blockade of intracellular thymidine synthesis, since 5FUDR itself was found not to be mutagenic, and 5FU-induced mutagenesis was not antagonised by supplying thymidine monophosphate (dTMP) to a dTMP permeable strain. In any case, observations of the molecular changes accompanying petite induction in log phase cells ruled out the possibility that mutagenesis resulted simply from the dilution out of replication blocked mitDNA molecules, since the appearance of mutants coincided with the synthesis of altered mitDNA molecules. In different strains, the resulting defective molecules were either maintained, giving rise to suppressive rho- petites, or completely degraded, to give pure clones of neutral rho0 mutants. It is suggested that this degradative process was a conseuqence of the incorporation of 5FU into RNA.

Centrifugation, Isopycnic↗
PubMed1980

Isolation of a single-stranded extrachromosomal DNA from germinating wheat embryos.

Newly-synthesized cytoplasmic non-mitochondrial DNA was isolated from wheat embryos which had been germinated in the presence of [14C]-thymidine for a time period not long enough to trigger the first post-dormant round of the nuclear DNA replication. This extrachromosomal DNA fraction consisted of linear single-stranded polydeoxyribonucleotide chains, corresponding in size to approximately 1.1 X 10(6) daltons, and amounted to about 0.5% of the total cellular DNA content. It is suggested that the appearance of the newly-synthesized polydeoxyribonucleotide chains in the cytoplasm may be a physiological signal for the initiation of the nuclear DNA replication in germinating wheat embryo cells.

Centrifugation, Density Gradient↗
PubMed1995

Parasitic apicomplexans harbor a chlorophyll a-D1 complex, the potential target for therapeutic triazines.

Ultrastructural evidence is presented for the presence of plastid-like organelles in Toxoplasma gondii, Sarcocystis muris, Babesia ovis, and Plasmodium falciparum. In addition, it was shown that merozoites of T. gondii contain protochlorophyllidae a and traces of chlorophyll a bound to the photosynthetic reaction centers I PS I and PS II. A psbA gene was isolated from merozoites of S. muris by the polymerase chain reaction (PCR). Partial sequencing of the PCR product revealed that the herbicide-binding region is highly conserved. Therefore, it is likely that the sensitivity of apicomplexans to the herbicide toltrazuril depends on the interaction of the herbicide with the D1 protein of the photosynthetic reaction center of the parasite's organelles.

Amino Acid Sequence↗
PubMed1977

Incompatibility and bacteriophage inhibition properties of N-1, a plasmid belonging to the H2 incompatibility group.

N-1, a plasmid isolated from a strain of Shigella flexneri in Japan more than 10 years ago, mediates the phage inhibition phenotype which has recently been found to be characteristic of plasmids of the H2 incompatibility group. Using the criteria of phage inhibition, surface exclusion and incompatibility, the N-1 plasmid is shown to be closely related to H2 plasmids isolated from non-typhoid salmonella and distantly related to H1 plasmids isolated from Salmonella typhi. Plasmids of other incompatibility groups did not show the H2 type of phage inhibition.

Antigens, Bacterial↗
PubMed1980

Population genetic analysis of human hand preference: evidence for generation differences, familial resemblance, and maternal effects.

Hand preference data were collected from 2818 subjects in 616 families using an adapted version of the Edinburgh Inventory. Population genetic analysis of these data and those from previous population studies, altogether encompassing 38,505 subjects in 8572 families, reveals evidence for generation differences, familial resemblance, and maternal effects.

Extrachromosomal Inheritance↗
PubMed1981

Maternal effects in mice: influence of parents' and offspring's genotypes.

Different backcrosses between the F1 and two inbred parental strains (C and B6) were compared for swimming speed in water escape task and for swimming ability in a water channel. Results showed maternal effects, mice from an F1 dam swimming faster than mice from an inbred dam. Albino mice appear to be more sensitive to these effects could also be related to the inbreeding level of the offspring, the reciprocal backcrosses showing differences, whereas the two reciprocal F1's did not. Maternal effects appear to be greater in the water escape task than in the water channel situation.

Animals↗
PubMed1993

Retrospective study of the parental origin of the extra chromosome in trisomy 18 (Edwards syndrome).

The parental origin of the extra chromosome in trisomy 18 was traced in 30 informative families using highly polymorphic (CA) repeats mapped on the long arm of chromosome 18. Proband DNA was recovered from slides of chromosome preparations in 28 cases and from paraffin-embedded tissues in two cases. The extra chromosome was found to be of maternal origin in 26 cases (86.7%), and paternal origin in 4 cases (13.3%).

Adult↗
PubMed1987

Correction of deletions in mammalian cells by gene conversion.

We have constructed substrates to study the conversion of deletions in mammalian cells both extrachromosomally and after the stable integration of the substrates into the chromosome. These substrates were designed to study gene conversion without the complication of reciprocal recombination events. The substrates contain insertion or deletion mutations of the neomycin resistance gene (neo) and an internal, homologous fragment of the neo gene (neo-526), such that gene conversion from neo-526 to the mutated neo gene restores a functional neo gene. We have shown that extrachromosomally insertions of 10 bp or deletions of 22 or 167 bp are converted to wild-type at similar frequencies (1-6 X 10(-4)). Chromosomal gene conversion occurred at frequencies of about 10(-6)-10(-7) per cell generation. As expected from the experimental design, all recombination events analyzed in mammalian cells using these substrates appear to be due to gene conversion.

Animals↗
PubMed1982

Chromosome-mediated transfer and amplification of an altered mouse dihydrofolate reductase gene.

We have conferred methotrexate resistance on mouse 3T6 fibroblasts by chromosome-mediated transfer of an altered dihydrofolate reductase gene encoding a highly methotrexate-insensitive enzyme. The methotrexate-resistant 3T6 cell line from which the chromosomes were prepared contains multiple copies of the altered dihydrofolate reductase gene, all of which appear to reside on double-minute chromosomes. Transformants selected at 0.2 microM methotrexate contain 10-20 times more of the transferred altered gene than of the resident normal gene. The altered genes are associated with double-minute chromosomes and are permanently lost following growth of the transformants in the absence of methotrexate. Growth of the transformants in increasing concentrations of methotrexate leads to the emergence of cells which have accumulated double-minute chromosomes and which have amplified only the transferred dihydrofolate reductase gene.

Animals↗
PubMed1978

Conservation of genes coding for proteins synthesized in human mitochondria.

Proteins synthesized in mitochondria of 27 different human cell lines, identified by labeling with [35S]methionine in the presence of cycloheximide, have been enumerated and their electrophoretic mobilities determined by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis and fluorography. Twelve bands were observed in all cell lines. In 24 cell lines, the electrophoretic mobilities of the proteins were the same regardless of race, sex, tissue of origin, cell type, viral transformation, or premature biological aging syndromes. The patterns obtained for the remaining cell lines, HeLa, KB, and Hep-2 were identical. These cell lines showed one protein component that was absent in the 24 others, and lacked a component present in these cell lines. Since it has been previously asserted that KB and Hep-2 are HeLa cells, the data indicate that one basic pattern exists in human cells with a variant of unknown origin occurring in HeLa cells.

Cell Line↗