An SLR1 promoter is a tissue-specific promoter for exocyst genes which is strongly expressed only in stigma and not in the pollen or style. Suppose that we have an RNA knockout gene construct as SLR1:SEC3a AS (along with an NOS terminator), where SEC3a is one of the subunits of the exocyst genes to be knocked-out in stigma. During RNA isolation after the gene knockout of SEC3a subunit in stigma, the use of tissues only from stigma is barely sufficient; hence incorporation of tissues from style becomes inevitable. This engenders the expression of SEC3a mRNA level even from style tissues despite of our stigma-centric SLR1 promoter in the knock-out construct during the qRT-PCR analysis. So if the relative expression of SEC3a mRNA during the real-time PCR analysis for the wild type plant is higher than that of these transgenic knockout lines, can we infer this to have espoused successful gene knockout of stigma SEC3a mRNA levels? Kindly clarify your position.
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