beta-Glucuronidase mRNA was purified from androgen-induced mouse kidney by immunoadsorption of polysomes to protein A-Sepharose. Cell-free translation of mRNA isolated from the protein A bound RNA followed by immunoprecipitation revealed that beta-glucuronidase mRNA represented approximately 2% of the purified mRNA fraction. This mRNA preparation was used to produce complementary DNA clones by recombination with pBR322. Clones containing sequences that were enriched during the purification procedure were selected by differential colony hybridization. These were further screened for homology with beta-glucuronidase mRNA by hybrid-selected translation. A beta-glucuronidase cDNA clone, designated pGUS7, was identified by these criteria. With this plasmid, the abundance of beta-glucuronidase mRNA in total poly(A) mRNA from androgen-induced mouse kidney was estimated to be less than 0.04%. The beta-glucuronidase cDNA plasmid hybridized to a mRNA of 2.6 kb in length, which was induced in an androgen receptor dependent fashion over a time course of 21 days. Treatment of female mice with a single dose of testosterone (10 mg) revealed that beta-glucuronidase mRNA concentration begins to increase between 12 and 24 h after hormone administration.
The lipopolysaccharides (LPSs) of five isolates of Pasteurella multocida from rabbits were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblots, and enzyme-linked immunosorbent assay. Silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of purified unaggregated LPSs resembled those of semirough strains of gram-negative enterobacteria and consisted of one or two bands that migrated within an interval just ahead or slightly behind the migration of the Ra chemotype of "Salmonella minnesota," which has a molecular size of 4.3 kilodaltons. Polyclonal rabbit antisera to P. multocida whole cells used in Western blots and enzyme-linked immunosorbent assays of unabsorbed and LPS-absorbed antisera revealed that the LPS of these isolates of P. multocida contained at least two types of antigens: a nonserospecific antigen and a serospecific antigen. The LPSs of four isolates each had a different serospecific antigen. The nonserospecific antigen was expressed in two isolates and was the only demonstrable LPS antigen in one other isolate.
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