model. To compare the translational efficiencies of normal and mutant TPO mRNAs, the DG mutation was introduced into the 58-UTR of TPO cDNA by recombinant polymerase chain reaction (PCR) and subcloned into the pcDNA3 vector, as previously described.2 Mutant and control TPO mRNAs were transcribed in vitro using T7 polymerase and translated in reticulocyte lysate in the presence of 35S-methionine...

Even a cursory review of the role of proteases in both muscle growth and in meat quality would require a lengthy treatise that could easily become too discursive to be either informative or easily understood. Besides, the role of proteases in meat quality (i.e., meat tenderness) has been clarified significantly during the last 15 years. A large amount of evidence has accumulated to indicate tha...

The 5’-flanking regions (promoters) of the bovine αs1-, αs2and β-casein genes were analysed for DNA sequence variants using PCR/RFLP in Polish Red (PR) and Black-and-White (BW) cattle. The polymorphic sites occurred at positions –1084 and –186 in the promoter region of the αs2-casein gene and at –728 and –109 in the αs1and β-casein genes, respectively. These polymorphic sites were located withi...

The in vitro pull-down assay is a well-established method to confirm direct binding in protein-protein interactions that was inferred from other interaction assays, such as two-hybrid analysis (1). The assay is usually carried out using glutathione-S-transferasetagged or His-tagged fusion proteins as the pull-down drivers and in vitro-translated 35S-labeled proteins as probes to detect interact...

The I region of the murine major histocompatibility complex (MHC) encodes a group of membrane alloantigens (Ia antigens) which appear intimately associated with immune response (Ir) genes mapped to this region (1, 2). Although five I subregions have been defined by serological and functional analysis (3), Ia antigens have been demonstrated by immunoprecipitation techniques for only the A and E ...

The standard enzyme-linked immunosorbent assay (ELISA) for anticardiolipin antibodies (ACA) detects a heterogenous group of antibodies against cardiolipin on its own, b2glycoprotein I (b2GPI), and, potentially, other phospholipidbinding plasma proteins from bovine or human origin. In an attempt to identify new proteic targets of ACA, we selected 6 patients who possessed cofactor-dependent ACA b...

The CWI of higher plants is a structurally well-characterized basic protein (pI approximately 9.5) showing a complex glycosylation pattern. CWIs of severa1 species have been characterized at the protein leve1 (Laurière et al., 1988; Weil and Rausch, 1994, and refs. cited therein). Recently, complete cDNA clones have been obtained for CWIs of carrot (Sturm and Chrispeels, 1990), Arabidopsis thal...

odium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) is the most widely used analytical method to resolve separate components of a protein mixture. It is almost obligatory to assess the purity of a protein through an electrophoretic method. SDS-PAGE simultaneously exploits differences in molecular size to resolve proteins differing by as little as 1% in their electrophoretic mobil...

Intact chloroplasts, purified from spinach leaves by sedimentation in density gradients of colloidal silica, incorporate labeled amino acids into at least 16 different polypeptides of the thylakoid membranes, using light as the only source of energy . The thylakoid products of chloroplast translation were visualized by subjecting membranes purified from chloroplasts labeled with [88S]methionine...

Using tracer methods, insulin stimulates muscle protein synthesis in vitro, an effect not seen in vivo with physiological insulin concentrations in adult animals or humans. To examine the action of physiological hyperinsulinemia on protein synthesis using a tracer-independent method in vivo and identify possible explanations for this discrepancy, we measured the phosphorylation of ribosomal pro...