The gene defective in the kar mutant codes for an endothelin-converting enzyme, Ece2, which activates endothelin ligands by proteolytic cleavage.
This means that mutants coding for maternal care have a greater probability of fixation than those coding for paternal care, even if the effect of maternal and paternal care on offspring survival is identical.
Briefly, MCF-7 cells were seeded on 12-well plates and transfected with 50 ng/well of plasmid pRLuc-Bax coding for BRET donor and increasing amounts (for saturation curves) or 200 ng (for ABT-737 dose response curves) of pS62A-eYFP-Bcl-xL or pS62D-eYFP-Bcl-xL mutants coding for BRET acceptor (or with corresponding pCMV-Bcl-xL for control).
To illustrate the many effects of reproductive variance on the evolution of dioecious species, we calculated the probability of fixation of mutants coding for maternal and paternal care for sons and daughters in a situation where the survival of sons is highly correlated within broods.
It is very difficult to see any apparent patterns of association between the original double-mutant fitness measurements, even for those mutant pairs coding for known genetic interaction.
For constructs used in the transient assays, full-length wild type or mutant coding sequences (CDS) of OVATE, SlOFP20, SlTRM3/4, SlTRM5 and SlTRM25 were cloned into pENTR/D-TOPO Gateway entry vector (Invitrogen, Carlsbad, CA) following the manufacturer's protocol.
A cDNA of mRNH coding mRI (with 456 amino acid residues) [ 14] and its PCR amplified spontaneous mutant (coding truncated product ΔmRI due to Glu340 was mutated to a stop code) were used for gene fusions.
To measure the efficiency of this step, we delivered [H]Phe-tRNAPhe to WT and mutant 80S IRES (coding for FVKM) shrimp ribosome complexes in the presence of eEF1A-GTP (which forms a ternary complex, TC, with ac-tRNA) and eEF2-GTP and collected these complexes by ultracentrifugation through a sucrose cushion (diagrammed in Figure 5 figure supplement 1).
Most times the mutant gene codes for a protein that boosts the formation of globs of protein debris.
