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Name:_______________ Quiz 7: Biol 203 Fall 2 aless-like (Arl) gene below carries a homeodomain DNA binding domain (HD). e, it has a proven activator domain (A) and repressor domain (R). A-R-HD reveal no function, however, cDNAs with A-HD have activator like and R-HD have repressor like properties. No cDNAs are found with HD alone. function mutation for this gene in the HD is has no toes in homozygous (homo) animals. have 5 toes, whereas heterozygous (het) loss of function animals have 4 toes. dentified several heterozygous mice that have 6 or more toes and you have identified a es a BamHI site in the gene, which correlates with the phenotype. Lab:______________ 24pts Coding exon 1 contributes 100 amino acids (10Kd) to the protein, exon 2 contributes 5Kd, exon 3 contributes 20Kd and exon 4 contributes 30Kd to the protein. he molecular weights (MWs) of the proteins derived from the WT gene?_3pts.__45kd, 60kd, 65kd_ d the SNP likely be in the gene?_2pt._ SNP in either intron or exon: Needs to affect A or R protein or splici this SNP mutation do to the eventual mRNA and/or protein?_2pt._stop codon, amino acid shift, makes say that it will create a null mutation or a loss of function mutation. ow you defined the SNP, what are the MWs of proteins produced in homo SNP animals? e three proteins specified in answer 1 will be lost, unless it is a missense mutation Arl do in the toe forming pathway? to say that it promotes toe formation or inhibits a protein that normally makes toes FLP analysis, describe what would be seen using the HD portion of the coding sequence dioactive hybridization probe after Southern blotting of digested genomic DNA for ous, heterozygous animals containing the SNP and WT animals. make sure to label your digests and genotypes for each lane of your blot, as the sizes of fragments produced after hybridization. A HD R EcoR1 EcoR1 10Kb EcoR1 fragment Arl Q1(12pts). Q2(12pts).

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Quiz 7: Biol 203 Fall 2013. The Arristaless-like (Arl) gene below carries a homeodomain DNA binding domain (HD). Furthermore, it has a proven activator domain (A) and repressor domain (R). cDNAs with A-R-HD reveal no function, however, cDNAs with A-HD have activator like - PowerPoint PPT Presentation

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Name:_______________Quiz 7: Biol 203 Fall 2013The Arristaless-like (Arl) gene below carries a homeodomain DNA binding domain (HD).

Furthermore, it has a proven activator domain (A) and repressor domain (R).cDNAs with A-R-HD reveal no function, however, cDNAs with A-HD have activator likeproperties and R-HD have repressor like properties. No cDNAs 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 SNPthat creates a BamHI site in the gene, which correlates with the phenotype.

Lab:_______________

24pts

Coding exon 1 contributes 100 amino acids (10Kd) to the protein, exon 2 contributes 5Kd,exon 3 contributes 20Kd and exon 4 contributes 30Kd to the protein.

What are the molecular weights (MWs) of the proteins derived from the WT gene?_3pts.__45kd, 60kd, 65kd_

Where would the SNP likely be in the gene?_2pt._ 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 mRNA and/or protein?_2pt._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? 2pts. 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? 3pts they have to say that it promotes toe formation or inhibits a protein that normally makes toes

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.

A HD R

EcoR1 EcoR1

10Kb EcoR1 fragment

Arl

Q1(12pts).

Q2(12pts).