Question

May you check if this answer is right and add any missing information to this answer (for example the origin of replication, etc.)

Review of Concepts: Gene Identification and Gene Knockout You want to study the physiological role of a gene you just cloned by making a "Knock-out" mutant mouse. Your gene has 5 exons (see below) and you want to mutate it by inserting neomycin resistance gene (neo) into exon 3 1 2 probe 3 4 Eco RI Eco R1 Eco Ri

1) Using the map above, draw a picture of an appropriate targeting vector and draw a map of the gene after neo has been inserted.

Answer 1

Your answer is correct however the third exon needs to flank the insertion region atleast on one side or both the sides and the fourth exon needs to be labelled. Otherwise its correct. Please refer the picture that I have attached, the third exon is intersected by the neo insertion. This will ensure a knockout without much workk on the existing DNA. Since direct deletion to replace the insertion of the neo gene cannot be done in 1 step.