Question

How are stable mutations insured in transposon mutagenesis?

Answer 1

During transposon mutagenesis, stable mutation are ensure by making sure that the transposase enzyme gene ( transposase cleaves the DNA at the target sites (inverted repeats flanking the transposon), generating 3' overhangs). producing enzyme which enables the transposon movement is not a part of the transposon region/ transposable element on the plasmid vector. The transposase can be expressed either on a separate plasmid, or on the plasmid containing the gene to be integrated. Alternatively, an injection of transposase mRNA into the host cell canalso be used to induce translation and expression. Therefore, the movement of transposon after its first insertion is hindered and the mutation becomes relatively stable.