Genetic mapping, cloning, and identification of <i>mfap-1</i>.

<p>(A) Genomic location of the <i>n4564 n5214</i> mutation based on genetic mapping using visible markers and SNPs (<i>e.g.</i>, SNP T01H8: 7110 on the left and SNP F36A2: 3334 on the right). Cosmids tested in rescue experiments are labeled. Only cosmid <i>F43G9</i> rescued the three phenotypic characteristics of <i>mfap-(n4564 n5214)</i> individuals: suppression of the Unc phenotype of <i>unc-93(e1500)</i> animals; partial sterility at 20°C; and temperature-sensitive lethality at 25°C. (B) Transgene rescue experiments. Transgenes (<i>nEx</i>) were injected into the gonads of <i>mfap-1(n4564 n5214); unc-93(e1500)</i> animals, and lines stably transmitting the transgenes were established. Transgenic lines were analyzed for the rescue of each of three abnormalities: suppression of the Unc phenotype of <i>unc-93(e1500)</i> animals; partial sterility at 20°C; and temperature-sensitive lethality at 25°C. <i>mut</i>: the <i>n4564 n5214</i> mutation. (C) Gene structure of <i>mfap-1</i>. Black boxes: coding exons. Open boxes: 5′ and 3′ UTRs. Positions of start (ATG) and stop codons (TGA) are indicated. The sites of the <i>n4564 n5214</i> mutation, the <i>n5214</i> mutation and the <i>tm3456</i>Δ deletion are labeled. (D) Sequence alignment of predicted MFAP-1 proteins from <i>C. elegans</i>, <i>Drosophila</i>, chicken and human. *: amino acids mutated in <i>mfap-1(n4564 n5214)</i> mutants. Amino acids conserved in at least three orthologs are darkly shaded, while amino acids with similar physical properties or conserved in two orthologs are lightly shaded.