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Title: Production of mRNA from the cry1Ac Transgene differs among Bollgard® lines which correlates to the level of subsequent protein.

Author
item Adamczyk, John
item Perera, Omaththage
item Meredith Jr, William

Submitted to: Transgenic Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/11/2008
Publication Date: 7/2/2008
Citation: Adamczyk Jr, J.J., Perera, O.P., Meredith Jr, W.R. 2009. Production of mRNA from the cry1Ac Transgene differs among Bollgard® lines which correlates to the level of subsequent protein. Transgenic Research. 18:143-149.

Interpretive Summary: Clear differences in survival and development were observed when certain caterpillars were fed different cotton varieties containing an insect-controlling gene (Bt). For the first time, these cotton varieties were bred for different amounts of Bt. Insects that fed on the variety that contained the lower amount of Bt survived better and developed faster than those caterpillars fed the other variety which contain a higher amount of Bt. However, the plant-mechanism for which this occurs was previously unknown until the present study. Using a technique called quantitative real-time polymerase chain reaction (qPCR), we developed a method to determine if differences in the level of Bt protein could be correlated to the RNA transcripts. Our data shows that the RNA transcript differs among Bt varieties and are correlated with corresponding Bt protein levels.

Technical Abstract: Commercial cultivars of Bollgard® cotton, Gossypium hirsutum L., differ in the amount of expressed Cry1Ac protein. However, the plant-mechanism for which this occurs is still unknown. Using quantitative real-time polymerase chain reaction (qPCR), we developed a method to determine if differences in the overall level of Cry1Ac among Bollgard® lines could be correlated to the mRNA transcripts. Our data shows that the cry1Ac mRNA transcript differs among Bollgard® lines and are correlated with corresponding Cry1Ac protein levels. In addition, qPCR based methods can efficiently be employed to quantify Cry1Ac protein expression levels in transgenic cotton cultivars. We postulate that qPCR based methods could be successfully employed for quantifying expression levels of transgenes in plants carrying different Bt proteins.