A cryptic ribosome binding site, false signals in reporter systems and avoidance of protein translation chaos.

The expression of reporter gene may be induced by activation of cryptic signalling sequences, as we found while constructing the mutS-lacZ fusion gene. We cloned the Escherichia coli lacZ gene encoding beta-galactosidase into a plasmid vector carrying the Thermus thermophilus mutS gene. The clones expected to produce beta-galactosidase as the C-terminal fusion were selected for the complementation of beta-galactosidase activity in a lacZ deficient E.

Further characterization of the combining sites of Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin A(4).

Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin A(4)(GS I-A(4)), which is cytotoxic to the human colon cancer cell lines, is one of two lectin families derived from its seed extract. It contains only a homo-oligomer of subunit A, and is most specific for GalNAcalpha1-->. In order to elucidate the GS I-A(4)-glycoconjugate interactions in greater detail, the combining site of this lectin was further characterized by enzyme linked lectino-sorbent assay (ELLSA) and by inhibition of lectin-glycoprotein interactions.

Multi-antennary Gal beta1-->4GlcNAc and Gal beta1-->3GalNAc clusters as important ligands for a lectin isolated from the sponge Geodia cydonium.

The affinity of a lectin from the sponge Geodia cydonium (GCL-I) for multi-antennary Gal beta1-->4GlcNAc and Gal beta1-->3GalNAc ligands was studied by both the biotin/avidin-based microtiter plate lectin binding assay and the inhibition of lectin-glycoform interaction. Among the glycoforms tested for binding, GCL-I reacted strongly with three multi-antennary Gal beta1-->4GlcNAc clusters containing glycoproteins (asialo human and bovine alpha1-acid gps and asialo fetuin), T (Gal beta1-->3GalNAc) rich glycoprotein from porcine salivary gland, asialo bird nest gp, and human blood group A active cyst gp, while human and bovine alpha1-acid gps, fetuin, and Tn containing gps were inactive. Among the haptens tested for inhibition, tri-antennary Gal beta1-->4GlcNAc (Tri-II) was about 1500, 72, and 72 times more active than GalNAc, Gal beta1-->4GlcNAc (II), and Gal beta1-->3GalNAc (T), respectively.

Further characterization of the binding properties of a GalNAc specific lectin from Codium fragile subspecies tomentosoides.

Previous study on the binding properties of a lectin isolated from Codium fragile subspecies tomentosoides (CFT) indicates that this lectin recognizes the GalNAc alpha1--> sequence at both reducing and nonreducing ends. In this study, the carbohydrate specificity of CFT was further characterized by quantitative precipitin (QPA) and inhibition of lectin-enzyme binding assays. Of the glycoforms tested for QPA, all asialo-GalNAc alpha1--> containing glycoproteins reacted well with the lectin.