Question

6. A sequencing chromatograph just arrived! a) Label each peak with an appropriate base (see Olga) b) Highlight mutation(s) site c) Characterize each chromatograph based off mutation(s) you found (give the name(s) of the mutation(s), briefly explain how it would affect the protein. Who is healthy/carrier/affected, why? d) Most importantly: Which mutation did Alexey have in his F9 gene?! e) How did this affect his blood clotting Factor IX? To answer these questions successfully, use the following sequence of the full Exon 3. intron 3. Exon 4 and a part of intron 4 (coding strand). Hint: you are in frame 1 ACT|GAATTT etc. 5'ACTGAATTTTGGAAGCAGTATGTTGgtaagcacctatctcaaagATGGAGATCAGTGTGAGTCCAATCCATG TTTAAATGGCGGCAGTTGCAAGGATGACATTAATTCCTATGAATGTTGGTGTCCCTTTGGATTTGAAGGAAAGA ACTGTGAATTAGgtaagtaactattttttqaata 3' - coding strand Sample Chromatograph Explanation Alexandra wwwmim Sequence: Olga CTCAA AGATGG Tatiana Sequence: MWWW Maria Sequence: Anastasia Sequence: wimm Miam Alexey Sequence:

Answer 1

Allexandra sequence = CTCAGAGATGG

Olga = CTCAAAGATGG

Tatiana = CTCAAAGATGG

Maria = CTCAAAGACGG

Anastasia = CTCAGAGATGG

Alexey = CTCAGAGATGG

Alexandra :
ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.ca-G-.agA.TGG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
explanation : mutation in the intron 3 sequence . CAA to CAG both codes for same amino acid. Affected individual

Oliga :
ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.caa.agA.TGG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
Explanation: no mutation - normal

Tatiana
ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.caa.agA.TGG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
Explanation : no mutation - normal
Maria :
ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.caa.agA.-C-GG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
explanation: arginine to tryptophan mutation - affected
Anestesia :

ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.ca-G-.agA.TGG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
explanation : mutation in the intron 3 sequence . CAA to CAG both codes for same amino acid. Affected individuals
Alexey :

ACT.GAA.TTT.TGG.AAG.CAG.TAT.GTT.Ggt.aag.cac.cta.tct.ca-G-.agA.TGG.AGA.TCA.GTG.TGA.GTC.CAA.TCC.ATG. TTT.AAA.TGG.CGG.CAG.TTG.CAA.GGA.TGA.CAT.TAA.TTC.CTA.TGA.ATG.TTG.GTG.TCC.CTT.TGG.ATT.TGA.AGG.AAA.GAA.CTG.TGA.ATT.AGg.taa.gta.act.att.ttt.tga.ata. 3'
explanation : mutation in the intron 3 sequence . CAA to CAG both codes for same amino acid. Affected individual

D) mutation in the intron 3 sequence . CAA to CAG both codes for same amino acid

Homologous mutation

E) specifity of factor 9 may differ