The Arristaless-like (Arl) gene below carries a homeodomain DNA binding domain (HD). Quiz 7: Biol 203 Fall 2013 Furthermore, it has a proven activator domain (A) and repressor domain (R). Name: ________ c. DNAs with A-R-HD reveal no function, however, c. DNAs with A-HD have activator like Lab: ________ properties and R-HD have repressor like properties. No c. DNAs are found with HD alone. A loss of function mutation for this gene in the HD is has no toes in homozygous (homo) animals. WT animals have 5 toes, whereas heterozygous (het) loss of function animals have 4 toes. You have identified several heterozygous mice that have 6 or more toes and you have identified a SNP that creates a Bam. HI site in the gene, which correlates with the phenotype. 24 pts Arl A R HD 10 Kb Eco. R 1 fragment Eco. R 1 Coding exon 1 contributes 100 amino acids (10 Kd) to the protein, exon 2 contributes 5 Kd, exon 3 contributes 20 Kd and exon 4 contributes 30 Kd to the protein. Q 1(12 pts). What are the molecular weights (MWs) of the proteins derived from the WT gene? _3 pts. __45 kd, 60 kd, 65 kd_ Where would the SNP likely be in the gene? _2 pt. _ SNP in either intron or exon: Needs to affect A or R protein or splicing (not HD). What will this SNP mutation do to the eventual m. RNA and/or protein? _2 pt. _stop codon, amino acid shift, makes a mistake in splicing but they CANNOT say that it will create a null mutation or a loss of function mutation. Based on how you defined the SNP, what are the MWs of proteins produced in homo SNP animals? 2 pts. one of the three proteins specified in answer 1 will be lost, unless it is a missense mutation What does Arl do in the toe forming pathway? 3 pts they have to say that it promotes toe formation or inhibits a protein that normally makes toes Q 2(12 pts). Using RFLP analysis, describe what would be seen using the HD portion of the coding sequence as a radioactive hybridization probe after Southern blotting of digested genomic DNA for homozygous, heterozygous animals containing the SNP and WT animals. Please make sure to label your digests and genotypes for each lane of your blot, as well as the sizes of fragments produced after hybridization.
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