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Isaac Johnson
Isaac Johnson

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To analyze the entire aflatoxin cluster, including intergenic regions, Fig 4 shows the results of our comparative polymorphism analysis between the aflatoxin gene clusters of NRRL 3357 and AF70, for which we found 1192 transition or transversion mutations (1.78% of the aflatoxin gene cluster) that differentiated the two morphotypes. The cluster of NRRL 3357 alone contained 116 deletions (0.17%), while the cluster of AF70 contained 742 deletions (1.11%). In total, more than 2000 SNPs within the aflatoxin cluster alignment could be used to differentiate these strains based on genotype. The greatest span of cluster alignment (9825 bp) having the lowest percentage of SNPs (0.35%) was found to encompass the start of the aflC/aflD intergenic region through the end of the aflA/aflB intergenic region. The second greatest span and second lowest percentage (7146 bp and 0.39%, respectively) encompassed the aflR gene through the end of the aflJ/aflE intergenic region. The intergenic region with the highest percentage (14.15%) of SNPs per span of genomic sequence (1039 bp) was aflL/aflI, and the gene with the highest percentage of SNPs (9.06%) per span of genomic sequence (1335 bp) was aflO. The highest concentration of deletions for both strains was found to reside in the aflF/aflU (norB/cypA) regions of their aflatoxin clusters. This region in L-morphotype NRRL 3357 exhibited 34 deleted bases not observed in AF70, and the S-morphotype AF70 exhibited 619 deleted bases not observed in NRRL 3357. Since both isolates are of the same mating type (MAT1-1), the presence of SNPs would be required to help differentiate the sequences, but their respective sequences were identical so our MLST analysis was based on five genomic loci.




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The genes are shown as arrows and the intergenic regions are shown as boxes (A and B). Shading for each gene and intergenic region relates to the quantity of transition and transversion SNPs observed (legend). Any number noted above or below a gene or intergenic region for panel A represents the quantity of base pair deletions (bpd) found within the NRRL 3357 or AF70 cluster sequences, respectively. The boxed aflF/aflU regions in panel A are enlarged in panel B, for which the genes in this region (for both A. flavus morphotypes) are compared to the same (complete) genomic region in the SU-1 A. parasiticus strain. Areas noted with bpd indicate large-scale deletions observed.


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Closing Statement: The entity's displayed behaviors in this interview have been found to be contradictory to those recorded of Frédéric Chopin. However, other interactions with SCP-3357-1 have proven it capable of mimicking personalities and mannerisms accurately enough to be indistinguishable. 041b061a72


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