Isolation of DNA-free RNA, DNA, and proteins by cesium trifluoroacetate centrifugation.

The ability to simultaneously isolate intact DNA-free RNA, genomic DNA, and proteins from a biological specimen can be useful in cloning genes and analyzing gene expression. Equilibrium density gradient centrifugation with CsCl is a useful tool for fractionating, quantitatively separating, and characterizing RNA, DNA, and the total quota of proteins, respectively, based on differences in their buoyant densities. In the present study we have reexamined the rarely used cesium salt, cesium trifluoroacetate, for the same purpose.

Expression and characterization of human interferon-beta1 in the methylotrophic yeast Pichia pastoris.

We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHuIFN-beta1) was secreted from shake-flask-grown P. pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.

Induced synthesis of albumin-like protein in damaged rat reticulocytes.

In this study, we present evidence that red blood cell (RBC) membrane p68 in the Belgrade (b/b) rat is similar if not identical to rat serum albumin (RSA). Structural homology between RSA and the RBC p68 has been determined by a variety of biochemical and immunological criteria. This albumin-like protein is a normal constituent of rat RBC and it is partially exported by exosomes during erythroid differentiation.

The rat beta (b miny)-globin promoter: nuclear protein factors and erythroid-specific induction of transcription.

We show that the rat adult beta (b miny)-globin gene is transcriptionally active. The - 100-bp promoter region contains control elements that are important for the induction of transcription in rat erythroleukemia (REL) cells. By using DNaseI footprinting and gel mobility shift assays, we have shown that the CCAAT box, a regulatory element from an analyzed promoter region, binds NF-Y and GATA-1 transcription factors.

Differences in rat rbc cytosol induced after in vivo phenylhydrazine treatment.

Analysis of rat red blood cell (RBC) cytosol protein fraction after in vivo phenylhydrazine treatment revealed increased amounts of 68-kDa protein. This protein is present in trace amounts in normal rat RBC cytosol. We also present data that 68-kDa protein from RBC cytosol has an identical isoelectric point, partial proteolytic map and immunological determinants as a protein of the same molecular mass from rat exosomes.