Hello all,

we are working on functional validation of a transposon mutated virulence genes in Agrobacterium tumefaciens. However, in the mutant, when the gene expression is analyzed by qPCR, we are observing almost similar CT values for the mutated gene in mutants compared to control strain.

We have confirmed the mutants by sequencing and we are sure of the presence of the transposon sequence within gene and we assume that only a truncated protein is synthesized as it encounters a stop codon within transposon sequence. We have designed primers downstream of transposon insertion site.

Is it possible that the promoter transcribes some random gene sequences downstream of transposon as this operon is bicistonic and hence we are seeing this expression?

Thank you in advance for your help!