A major QTL controls susceptibility to spinal curvature in the curveback guppy.

Background: Understanding the genetic basis of heritable spinal curvature would benefit medicine and aquaculture. Heritable spinal curvature among otherwise healthy children (i.e.

Dominant marker vectors for selecting yeast mating products.

We have developed plasmid vectors to enable selection of diploids from mating reactions between haploid strains that lack compatible recessive genetic markers. The plasmids bear one of five different dominant selectable markers, kanMX4, hphMX4, natMX4, patMX3 or ZEO, a yeast origin of replication, and the URA3 gene. Diploids can be selected from mating reactions between haploids transformed with plasmids expressing different dominant markers, by using a combination of drugs that select for both markers.

Identification of protein interaction antagonists using the repressed transactivator two-hybrid system.

The repressed transactivator (RTA) yeast two-hybrid system was developed to enable genetic identification of interactions with transcriptional activator proteins. We have devised modifications of this system that enable its use in screening for inhibitors of protein interactions from small molecule compound libraries. We show that inhibition of protein interactions can be measured by monitoring growth in selective medium containing 3-aminotriazole (3-AT) and using this assay have identified inhibitors of four independent protein interactions in screens with a 23,000 small molecule compound library.

Disintegrator vectors for single-copy yeast chromosomal integration.

Vectors were developed for two-step chromosomal integration of reporter genes or expression constructs. With these vectors, integration produces a disruption of the ADE8, LYS2, MET15, LEU2, HIS3 or FCY1 genes, and integrants can be easily identified by replica-plating on selective media. Integration using these 'disintegrator' vectors produces a single-copy integration of the construct of interest at the junction of the marker deletion, and removes the additional plasmid sequences.

Overexpression of L-isoaspartate O-methyltransferase in Escherichia coli increases heat shock survival by a mechanism independent of methyltransferase activity.

Over time and under stressing conditions proteins are susceptible to a variety of spontaneous covalent modifications. One of the more commonly occurring types of protein damage is deamidation; the conversion of asparagines into aspartyls and isoaspartyls. The physiological significance of isoaspartyl formation is emphasized by the presence of the conserved enzyme L-isoaspartyl O-methyltransferase (PIMT), whose physiological function appears to be in preventing the accumulation of deamidated proteins.