This finding suggests that results obtained with Put3 have to be interpreted with caution. However, on the basis of our extended sequence-to-structure rules it should be possible to optimize Put3 for studies of transmembrane receptors. Angiopoietins are the ligands of Tie2 and have potential therapeutic applications in angiogenesis and prevention of vascular leakage. However, the large-scale production of recombinant angiopoietins, in particular angiopoietin-1, is hindered by the aggregation and insolubility of the proteins. We and others have produced soluble angiopoietin-1 variants by replacing the native oligomerization domains by coiled-coil domains from other proteins. Our designed trimeric variant using the coiled-coil domain of matrilin-1 as a fusion partner formed a mixture of trimers and tetramers and phosphorylated Tie2 to the same extent as the native protein. However, the minimal oligomerization state of angiopoietin-1 required for Tie2 activation is currently unknown. The Cort-Ir-M1-short sequence appears to be an excellent candidate fusion partner to address this question. Integrins comprise a large family of heterodimeric adhesion receptors consisting of a and b subunits.qPCR performance parameters are important in determining whether the target PCR product successfully doubles with each amplification cycle. To test for potential biases during the DNA extraction and PCR amplification, we compared two standard sets made from PrepMan extracts: the first quantified by direct zoospore counts and the second based on equimolar DNA concentration dilutions for each strain. If these two standard sets vary in amplification rates or in performance parameters among different Bd strains, we can infer that either the zoospore extraction efficiencies differ between the strains, or that genomic changes among strains cause differences in their amplification efficiencies. Using analyses of covariance, we tested for the effect of strains in the relationship between the number of genomic equivalents and Ct by fitting separate linear regressions for each standard set. If amplification efficiency among strains varied, we expected significantly different slopes and significant interaction terms across strains. If the number of ITS1 regions varied among strains, we expected differences in the y-intercept of the regression lines. We applied Tukey’s Honestly Significant Differences to determine which pairwise strain comparisons were statistically different. To quantify the number of ITS1/5.8S regions in each strain, we used a standard curve based on the ITS1 PCR-amplicon against each of the zoospore-based serial dilutions. We performed all statistical analyses using R. We compared the number of ITS1 haplotypes recovered by cloning and whole genome sequencing using a paired t-test. Finally, we calculated ITS1 haplotype frequencies for both data sets and estimated pairwise genetic differentiation among the ITS1 haplotypes for each Bd strain using FST statistics implemented in Arlequin 3.5.