A deficiency in factor Vill causes hemophilia A, a blood disorder. Researchers s

Question

A deficiency in factor Vill causes hemophilia A, a blood disorder. Researchers studying hemophilia A evaluated the DNA from an affected patient and the patient's unaffected mother. They analyzed the 186-kb-long factor VIII gene that includes 26 exons (See Chapter 14 in the textbook). After digesting the genomic DNA with Kpnl or Sstl and separating the products by gel electrophoresis, the researchers probed the DNA with a radiolabeled cDNA probe that binds the factor VIII gene in a region that includes exons 14-26. The size of the fragments and the corresponding exon (s) are shown on the right of autoradiograms. The researchers conclude that a transposon inserted into one of the exons of the factor Vill. E kb exon kb exon 30.0 (15-22) 19.1 (14) 21.0 (23-25) 12.7 (15-22) 8.3 (23-25) 7.3 (14) -5.5 (14) 5.3 (14) -4.3 (14) -4.0 (26) 3.2 (14) lane 1 lane 1 Sstl Kpnl

Explanation / Answer

Part B: In the KpnI digest, the patient has a transposon inserted in the exon 14 which is around 2.3Kb in length. This can be differentiated from the mother as she lacks both 5.3 and 4.3Kb fragment. But the patent has both these fragments while missing 7.3Kb fragment from the autoradiogram.

Part C: The hypothesis would be that there is an insertion of transposon in the exon 14 which gives different SstI and KpnI. The digestion pattern with SstI and KpnI is different between patient and the mother.

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