Proteolytic degradation of heterologous proteins expressed inAspergillus niger

Summary Proteolytic degradation of heterologous proteins expressed in the filamentous fungusAspergillus niger reduces the yield of authentic target protein. The activities ofA. niger proteases are differentiated by their effects on two proteins expressed and secreted fromA. niger: hen egg-white lysozyme and porcine pancreatic phospholipase A₂.     

Promoter methylation and progressive transgene inactivation inArabidopsis

Agrobacterium-transformedArabidopsis plants were generated and the stability of their T-DNA-encoded resistance to kanamycin was examined. Of seven families, each homozygous for a single insertion event, two showed progressive inactivation of resistance over four generations of inbreeding. Loss of resistance was associated with methylation of anSst II site in thenos promoter of the kanamycin resistance gene. Treatment of plant roots from inactive lines with the demethylating agent 5-azacytidine restored the ability of such lines to form callus on kanamycin-containing media. These observations are consistent with the view that methylation is a factor in the progressive inactivation of transgenes inArabidopsis.     

Vanadium-containing tunicate blood cells are not highly acidic

The intracellular pH of intact blood cells of the tunicate Ascidia nigra was measured by transmembrane equilibration of [¹⁴C]methylamine. The pH of unfractionated blood cells is 7.39±0.10. The pH of vanadocytes, determined in a fractionation study, is 7.2. Previously used methods, in which pH values less than 3.0 are inferred from cell lysis or vital staining experiments, are shown to be unsuitable for intracellular pH determination due to the chemical composition of these vanadium-containing cells.     

Comparison of content with synthesis,uptake and breakdown of GABA in substantia nigra of unoperated,hemitransected and kainate-treated rats

The synthesis, high affinity uptake and degradation of GABA were compared with GABA content measured simultaneously in the nigras of unoperated, hemitransected and kainate-treated rats. In intact animals nigral GABA concentration was linearly related to glutamate decarboxylase activity, but not to GABA aminotransferase (GABA-T) or GABA uptake. One week after hemitransection all three parameters showed a linear reduction with GABA content, suggesting striatonigral GABA neurones may account for as much as 90% of the GABA and GAD, 31% of the GABA-T and 58% of GABA uptake sites in whole nigra. Nigral kainate treatment unexpectedly reduced all four parameters in parallel by as much as 50%, consistent with the formation of an unselective lesion.