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At least 667 records · Page 37Linked to original sources
PubMed1995

Types of Listeria monocytogenes predicted by the positions of EcoRI cleavage sites relative to ribosomal RNA sequences.

By using taxonomic characters derived from EcoRI restriction endonuclease digestion of genomic DNA and hybridization with a labeled rRNA operon from Escherichia coli, a polymorphic structure of Listeria monocytogenes, characterized by fragments with different frequencies of occurrence, was observed. This structure was expanded by creating predicted patterns through a recursive process of observation, expectation, prediction, and assessment of completeness. This process was applied, in turn, to normalized strain patterns, fragment bands, and positions of EcoRI recognition sites relative to rRNA regions. Analysis of 1346 strains provided observed patterns, fragment sizes, and their frequencies of occurrence in the patterns. Fragment size statistics led to the creation of unobserved combinations of bands, predicted pattern types. The observed fragment bands revealed positions of EcoRI sites relative to rRNA sequences. Each EcoRI site had a frequency of occurrence, and unobserved fragment sizes were postulated on the basis of knowing the restriction site locations. The result of the recursion process applied to the components of the strain data was an extended classification with observed and predicted members.

Base Sequence↗
PubMed1980

The messenger RNA sequences in human fibroblast cells induced with poly rI.rC to produce interferon.

Induction of human fibroblast cells with poly rI.rC induces interferon mRNA which can be translated into interferon precursor in wheat germ cell free system or in Xenopus oocytes into biologically active interferon. The extent of gene expression in the poly rI.rC induced cells was compared to that of the uninduced cells by hybridization of the mRNA to complementary DNA. Homologous template driven hybridization of cDNA revealed the presence of two clearly defined transitions in the total poly A RNA from the induced cells; abundant class and a scarce class comprising approximately 37,000 diverse species of RNA. Heterologus hybridization of the cDNA with total uninduced mRNA showed that the majority of the mRNA sequences are the same in both the induced and uninduced cells. The results of the hybridization using cDNA prepared to the fraction enriched for interferon mRNA, however, showed that about 4% of the sequences present in the interferon enriched fraction are not present in the uninduced cells. These differences may result from the poly rI.rC induced alterations in gene expression.

Fibroblasts↗
PubMed1982

Abundant messenger RNA sequences common to transplanted mouse tumours of different tissue origins.

Radioactive cDNA, cell-free-synthesised on poly(A)-rich mRNAs from mouse FLS-ascites tumour cells, were separated into two fractions corresponding to abundant mRNA species and to rare mRNA species respectively, the former being nearly 100-times more abundant per cell than the latter. Aliquots of each fraction were hybridized to a large excess of RNA of different origins; three ascites tumours (the FLS, Krebs 2 and Ehrlich tumours) all transplanted in animals; one established line of Friend erythroleukemia cells serially passaged in culture; adult organs and whole mouse embryos. It was shown that all or nearly all the mRNA species, in particular the abundant ones, were present and similarly distributed in the three transplanted tumour strains studied, despite their different tissue origins. The great majority of abundant mRNA species were also present in Friend erythroleukemia cells but about two-times less frequent relative to transplanted tumours. Much greater differences were found as compared with non-malignant cells. In particular, although the majority of the abundant mRNA species were also found in all normal tissues studied, they were on the average 7-8-times less abundant in adult liver or in 10-days-old embryos, and 10-times less abundant in adult brain, than in tumour cells. Finally about 10% of the abundant RNA species common to all transplanted tumours appeared to be absent in normal cells, whatever their growth rates or stage of differentiation. The possible implications of these findings are discussed.

Animals↗
PubMedDate not supplied

Bead-based sandwich hybridization characteristics of oligonucleotide-alkaline phosphatase conjugates and their potential for quantitating target RNA sequences.

The hybridization characteristics of oligonucleotide-alkaline phosphatase conjugate probes were examined in bead-based sandwich hybridization reactions using single-stranded nucleic acid targets and oligonucleotide-polystyrene capture beads. Enzymatic activity was monitored using a chemiluminescent substrate and calibration plots of chemiluminescent signal versus conjugate concentration were used to estimate the sandwich hybridization efficiencies. Improved hybridization behavior was noted using glycerol as an additive and by increasing the length of the probe and alkyl spacer of the conjugates. The chemiluminescent assay is at least as sensitive as those employing 32P-labeled probes and can detect as little as 10-20 amol of target RNA. The linear relationship of chemiluminescent signal versus target assayed provides a method for quantitating unknown target samples. A single human immunodeficiency virus type 1 infected cell in a background of 10(6) uninfected cells is facilely detected when this enzyme-based detection assay is prefaced with a self-sustained sequence-replication amplification reaction.

Alkaline Phosphatase↗