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Submitted to: National Cotton Council Beltwide Cotton Conference
Publication Type: Proceedings Publication Acceptance Date: 1/30/2004 Publication Date: 6/1/2004 Citation: Zhu, Y., Adamczyk Jr., J.J. 2004. PCR confirmation of the cry1Ac gene in transgenic Bt (bollgard) cotton. Proc. Beltwide Cotton Prod. Res. Conf. CD ROM Interpretive Summary: To evaluate the purity of Bt cotton seed, we developed a polymerase chain reaction (PCR) technique to examine whether the Bt gene is present in transgenic cotton. PCR amplified a 1061 nucleotide DNA fragment that corresponded to represent the Bt gene inserted in cotton genome. A total of 150 cotton plants were tested, and all plants were confirmed to contain the Bt gene using this PCR technique. This technique will be used with ELISA to study Bt gene expression and Bt protein expression under different environmental conditions. Technical Abstract: Transgenic cotton (Gossypium hirsutum L.) with Bacillus thuringiensis Berliner -endotoxin genes has been widely adopted for suppression of lepidopteran pests. To account for inconsistency of the endotoxin gene expression, we developed a polymerase chain reaction (PCR) for checking the purity of transgenic cotton plants. A total of eight Cry1Ac genes were aligned for PCR primer design. A DNA fragment was amplified from Bt cotton, sequenced, and confirmed to be a portion of the Bt gene. A total of 150 cotton plants representing five cultivars were examined for the presence of the Bt gene. Results demonstrated that all of these cotton plants harbored the Bt endotoxin gene. This PCR technique can be used for future studies involving the Bt gene and protein expression. |
