Genetics: From Genes to Genomes
Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Chapter 14, Problem 30P

The sequence at one end of one strand of the Drosophila transposon Mariner is shown below (dots indicate sequences within the transposon):

5′ TTAGTTTGGCAAATATCTCCCTTCCGCCTTTTTGATCTTATGT... 3′

You obtain a mutant bacterial strain tagged with an engineered Mariner transposon, cut the genomic DNA from this strain with the restriction enzyme MboI (whose recognition site is ^GATC), and circularize the resultant DNA fragments by diluting the restriction enzyme digest and adding DNA ligase.

a. Design two 17 bp PCR primers that you could use to identify (by inverse PCR) the gene into which the transposon inserted.
b. What DNA sequence will be amplified from the circularized fragments of the mutant genome? Show the extent of this DNA sequence on a map of the genome of the mutant strain, indicating the locations of the transposon insertion and any relevant sites for the enzyme MboI.
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