On the processing of the prehormone of human placental lactogen (hPL) in the wheat germ cell-free system [proceedings].
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Biomedical subjects
Publications and source records attributed to J Mous.
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Oocytes from Xenopus laevis were injected with polysomes from normal human term placenta. Synthesis of the protein hormone Human Placental Lactogen (HPL) in the oocytes was demonstrated by specific immunoprecipitation with anti-HPL serum. Analysis of the immunoprecipitates on SDS-polyacrylamide gel revealed one peak, with a migration distance corresponding exactly to that of [14C]-Radioacetylated HPL added as a marker. Two days after injection the mRNA was still able to direct the synthesis of HPL.
Total polysomes from human term placenta were incubated in a wheat germ cell-free system during 1 hr at 25 degrees C. Human placental lactogen hormone was identified among the proteins synthesized in vitro by immune precipitation and subsequent sodium dodecylsulphate electrophoresis of the immunoprecipitate. The zone containing the radioactivity from the immunoprecipitate [3H]-labelled lactogen hormone comigrated exactly with the radioactivity zone from added [14C]-labelled marker hormone. This result indicates that the molecular weight of the synthesized product must be equal or very similar to that of the native protein hormone.
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A quantitative PCR assay for the detection of HIV-1 nucleic acids is described. The assay is based on a competitive internal standard nucleic acid which can be discriminated from target sequences by the presence of a new restriction enzyme site. The method was used to quantitate plasmid molecules containing HIV-1 sequences, HIV-1 DNA and HIV-1 RNA purified from HIV-1-infected tissue culture cells as well as HIV-1 DNA present in the peripheral blood mononuclear cells of an AIDS patient. The assay will be valuable for assessing viral load in AIDS patients and for the study of viral gene expression.