By John N.Abelson, Melvin I.Simon
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Transposon insertions can be used to construct deletions or duplications with defined endpoints, or can serve as sites of integration of other genetic elements. 8. When used as a recipient in a genetic cross, transposon insertions behave as point mutations in fine‐structure genetic mapping. 9. Transposon insertions can be obtained that are near but not within a gene of interest. Such insertions are useful for constructing defined deletions and duplications, as well as for genetic mapping. 10. Special transposons can be used to construct operon or gene fusions.
3. Start fresh 1‐ml overnight cultures of the S. enterica nadB and his auxotrophs in LB. Grow overnight with aeration at 37 . 6 ml of cells. 2 ml of the mixture onto each of 5 L‐Tc plates. Incubate overnight at 37 . 4. Replica print to E‐Tc plates (minimal E glucose), and incubate overnight at 37 . 5% of the TcR recombinants to be linked to either auxotrophic marker, and transduce the recipients to prototrophy. These will grow on the E‐Tc plates. 5. , 1996). 6. Prepare P22‐transducing lysates on each insertion mutant.
18, 97–100. [4] Identification of Essential Genes in Bacteria By DAVID R. HILLYARD and MICHAEL J. REDD Abstract Essential genes are identified in duplicated regions of the bacterial chromosome. Transposition of a vector that forms operon fusions into a strain carrying a chromosomal duplication allows insertion of the transposon into essential genes because a second copy of the essential gene is present. When the duplication is allowed to segregate, only the segregant that carries the copy of the intact essential gene survives.