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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

RNA was extracted from purified tachyzoites of Toxoplasma gondii (RH strain) by sequential centrifugation in guanidine hydrochloride, urea, and lithium chloride. The subunits of the RNA were characterized by denaturing and non-denaturing electrophoresis in agarose gels. Poly(A)+-RNA, purified by oligo(dT)-cellulose affinity chromatography, was translated in a rabbit reticulocyte lysate assay and the products were immunoprecipitated with an experimentally infected mouse serum and a naturally infected human serum. After sodium dodecyl sulphate-polyacrylamide gel electrophoresis, fluorography of the polypeptides confirmed that the mRNA translated specific parasite antigens.
Mol Biochem Parasitol 1986 Mar
PMID:Characterization and in vitro translation of Toxoplasma gondii ribonucleic acid. 242 Nov 63

Poly(A)+ RNA was isolated from maize scutella of different stages of post-germinative development and translated in vitro in a rabbit reticulocyte translation system. Immunoprecipitation of the translation products with CAT-2-specific antibody was used to quantitate the relative levels of translatable CAT-2 mRNA at each stage. The results show a close correlation between the developmental profile of Cat2 gene expression and the profile of CAT-2 mRNA levels. Evidence that the levels of CAT-2 mRNA are regulated by a temporal regulatory gene (Car1) is presented and the possible mechanism(s) of this regulation discussed.
Mol Gen Genet 1986 Apr
PMID:Cat-2 gene expression. Developmental control of translatable CAT-2 mRNA levels in maize scutellum. 242 49

Poly(C, G) random copolymer templates direct the oligomerization of 2-Me-ImpG and 2-MeImpC, resulting in the production of a variety of oligo(G, C)s. The efficiency of monomer incorporation into newly synthesized oligomers is greater for 2-MeImpG than for 2-MeImpC, and decreases for both monomers as the guanine content of the template increases. The relatively low efficiency of oligomerization on guanine-rich templates is largely a consequence of intra- and intermolecular template self-structure. The problem of template self-structure is clearly a major obstacle to the development of a system of self-replicating polynucleotides. The distribution of oligomeric products can be characterized in detail using high-pressure liquid chromatography on an RPC-5 column. Oligomers are separated on the basis of chain length, base composition and phosphodiester-linkage isomerism. Oligomers up to about the 12-mer, with base composition Gn, Gn-1C and Gn-2C2, have been identified. The 3' to 5' regiospecificity of the products is high, particularly for oligomers with base composition Gn.
J Mol Biol 1986 Apr 05
PMID:Non-enzymic template-directed synthesis on RNA random copolymers. Poly(C, G) templates. 242 55

Past work has shown that B lymphocytes from Peyer's patches (PP) and from spleen differ from each other in the following ways: (1) PP are enriched for IgA-producing precursor cells while splenic B-cells are a rich source of IgM-precursor cells; and (2) PP are a poor source of antibody-secreting cells when compared to the spleen. The reasons for these differences are unclear. In this work B-cells have been examined expressing newly regenerated surface immunoglobulin heavy chains in murine PP and spleens for immunoglobulin isotype expression. Dual color immunofluorescence demonstrated higher levels of B-cells regenerating two surface isotypes in the PP (IgM+ IgG+ = 14.9%; IgM+ IgA+ = 9.4%) than in the spleen (IgM+ IgG+ = 0.4%; IgM+ IgA+ = 0.2%). Poly A+ RNA was purified from these B-cells and compared for immunoglobulin heavy chain isotype expression by DNA-excess slot blot hybridization. Splenic B-cells contained higher amounts (at least two-fold) of Ig heavy chain-specific mRNA per microgram of polysomal RNA than did PP B-cells. Peyer's patches B-cells demonstrated slightly lower mu:alpha ratios than splenic B-cells. In vitro translation of the RNAs suggested higher levels of translatable alpha-specific Poly A+ RNA in PP B-cells than in B-cells from MLN or spleen; furthermore, splenic B-cell RNA contained higher levels of translatable mu-RNA than did the other tissues examined. Northern blot analysis of RNA derived from these tissues identified major mu-, gamma 2b-, and alpha-hybridizing sequences, though PP-derived B-cell preparations were shown to be enriched for the membrane forms of mRNA for gamma 2b and alpha when compared to the spleen-derived B-cell preparations. These results suggest that the level of differentiation of PP B-cells (that are capable of regeneration of surface Ig) may differ significantly from that of splenic B-cells.
Mol Immunol 1986 Jun
PMID:Comparison of immunoglobulin heavy chain isotype expression in Peyer's patch and splenic B-cells. 242 36

Total cellular Poly A+ RNA from TEPC15 myeloma and murine lymphoid tissues was analyzed by denaturing agarose gel electrophoresis and Northern blot hybridization to specific heavy chain constant region cDNAs; mu, gamma 2b and alpha RNA species were identified in each of the tissues and in the IgA producing TEPC15 myeloma. In total Poly A+ RNA from TEPC15 myeloma, alpha-cDNA hybridized predominantly to a 2.3 kb RNA species; 11.5 and 4.1 kb RNA species were evident as well. Successive hybridization of the same RNA to mu- and gamma 2b-specific cDNAs demonstrated the presence of both mu and gamma 2b specific RNA species with electrophoretic mobilities apparently identical to the 11.5 and 2.3 kb RNA species identified by alpha-specific hybridization. These data establish the presence of Poly A+ RNA species containing alpha, mu, and gamma 2b sequences in TEPC15 cells and suggest the transcription of RNA from both CH-containing chromosomes in TEPC15 myeloma (one chromosome in the TEPC15 cell line contains all the CH genes while the other chromosome has deleted all CH genes except alpha). In total Poly A+ RNA from normal mouse tissues (Peyer's patch and spleen) all three cDNA probes hybridized predominantly to an 11.5 kb RNA species. Primer extension experiments demonstrated that alpha cDNA could prime for the synthesis by reverse transcriptase of gamma 2b DNA when Peyer's patch Poly A+ RNA was used as the template. This suggests the existence of a single transcript containing alpha and gamma 2b sequences. Murine lymphoid tissues contained putative mRNAs for mu, gamma 2b and alpha heavy chain proteins, whereas TEPC15 myeloma polysomal Poly A+ RNA contained only alpha mRNA.
Mol Immunol 1986 Jun
PMID:Identification of the major immunoglobulin heavy chain poly A+ RNA in murine lymphoid tissue. 242 42

RNA has been prepared from promastigotes of Leishmania tropica and Leishmania d.donovani using three different methods. Extraction by hot phenol/isothiocyanate gave the best quantitative and qualitative results. The analysis of total RNA on methyl mercuric agarose gels shows that the large rRNA species is nicked: it is composed of a 630 and a 560 kDa molecule. The small rRNA species has a molecular weight of 800,000. Poly(A+) RNA can be translated in a rabbit reticulocyte lysate system. The newly synthesized products comprise high molecular weight proteins and show different patterns using RNA from L. tropica or from L. d. donovani promastigotes.
Mol Biochem Parasitol 1987 May
PMID:Characterization of RNA from Leishmania tropica and Leishmania d.donovani promastigotes. 244 Dec 55

Poly(A)+ RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences specific for nodules were selected by differential colony hybridization using 32P-labeled cDNA synthesized either from nodule poly(A)+ RNA or from poly(A)+ RNA of uninfected root as probes. Among the recombinant plasmids, the cDNA gene for leghemoglobin was identified. The protein structure derived from its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+ RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules.
Mol Biol Rep 1987
PMID:Molecular cloning of lupin leghemoglobin cDNA. 244 1

Poly(A)+ RNA was isolated from Ascaris suum body wall muscle and translated in a cell-free rabbit reticulocyte lysate system. Specific antisera and immunoglobulins against the alpha-pyruvate dehydrogenase and dihydrolipoyl transacetylase components of ascarid pyruvate dehydrogenase complex were used to immunoprecipitate individual radiolabelled polypeptides from the in vitro translation mixtures. Both polypeptides appeared to be synthesized as preproteins about 1.5 and 8 kDa larger than the corresponding native proteins. Incubation of the dihydrolipoyl transacetylase preprotein with an ascarid high-speed mitochondrial supernatant fraction resulted in the formation of a polypeptide with apparent molecular weight intermediate in size between the preprotein and the native enzyme. This processing was insensitive to phenylmethylsulfonyl fluoride and leupeptin but was completely abolished by EDTA. These results suggest that in A.suum, as in other organisms, mitochondrial matrix proteins coded by the nuclear genome are synthesized as larger preproteins and processed by a specific, metal-dependent mitochondrial matrix protease.
Mol Biochem Parasitol 1988 May
PMID:In vitro synthesis and processing of components of the Ascaris suum pyruvate dehydrogenase complex. 245 26

Poly(C,A) random copolymer templates direct the oligomerization of 2-MeImpG (2-MeImpX is the 5'-phospho-2-methylimidazolide of the nucleoside X) and 2-MeImpU, resulting in the production of a variety of oligo (G,U)s. This reaction is less efficient than comparable reactions involving poly(C,U) or poly(C,G) templates. The efficiency of monomer incorporation into newly synthesized oligomers is lower for 2-MeImpU than 2-MeImpG, and cannot be improved by increasing the concentration of 2-MeImpU relative to 2-MeImpG. This suggests that RNA templates containing runs of consecutive adenine residues would not be suitable for use in a chemical self-replicating system. The distribution of oligomeric products can be characterized in detail using high-pressure liquid chromatography on an RPC-5 column. Oligomers are separated on the basis of chain length, base composition, and phosphodiester-linkage isomerism. Oligomers up to about the 13-mer, with base composition Gn, Gn-1, U, and Gn-2, U2, have been identified.
J Mol Biol 1988 Aug 05
PMID:Non-enzymatic template-directed synthesis on RNA random copolymers. Poly(C,A) templates. 245 95

To study the role of a template sugar-phosphate backbone in the ribosomal decoding process, poly(U), poly(dT) and poly(dU)-directed cell-free amino acid incorporation was investigated under the influence of neomycin and high concentrations of Mg2+. The specificity of a factor-dependent translation system of Escherichia coli was shown to change according to the principle: "either ribo- or deoxyribopolynucleotide messenger". Poly(dT) is shown to be effectively translated in the absence of elongation factors, both at low (2 degrees C) and high (37 degrees C) temperature. Neomycin inhibits factor-free poly(dT) translation. Little or no poly(U) translation is observed in this system. A chromatographic analysis of the oligophenylalanine residues synthesized seems to show that translocation is the main step responsible for ribosome specificity to the ribo- or deoxyribopolynucleotide template in both factor-dependent and factor-free translation systems.
J Mol Biol 1988 Oct 20
PMID:The role of a template sugar-phosphate backbone in the ribosomal decoding mechanism. Comparative study of poly(U) and poly(dT) template activity. 246 69


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