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目的 检测转sck基因水稻T6代种子中外源蛋白CpTI及HPT的表达量。方法 对转基因水稻种子进行PCR检测;取外源基因PCR检测阳性植株的叶、茎杆、根部、种子等不同部位对CpTI和HPT蛋白表达进行Westernblot分析,并对PCR检测阳性植株的种子进行双夹心ELISA检测,以同种非转基因水稻M86种子作对照。结果 在转sck基因水稻T6代种子中扩增出sck和hpt基因;Westernblot结果显示转基因水稻的叶片、茎杆和根部的蛋白出现特异的杂交条带,种子蛋白中未出现特异杂交条带。ELISA检测结果显示,转基因水稻种子中CpTI蛋白低于双夹心ELISA的检测低限(<14 μg L) ,HPT蛋白没有被检出。结论 转基因水稻种子中CpTI蛋白含量低于检测低限,不能被定量,并且不含有可检测水平的HPT蛋白。
Objective To detect the expression of CpTI and HPT in T6 seeds of transgenic sck rice. Methods PCR and Western blot were used to detect the expression of CpTI and HPT protein in the leaves of transgenic rice by PCR with PCR, the leaves, stems, roots, seeds and other parts of the positive plants were detected by PCR. ELISA test to the same non-transgenic rice M86 seeds as a control. Results The sck and hpt genes were amplified from T6 seed of transgenic sck rice. Western blot showed that there were specific hybridization bands in the leaves, stems and roots of transgenic rice and no specific hybridization bands in the seed protein. ELISA results showed that the CpTI protein in transgenic rice was lower than the detection limit (<14 μg L) of double-sandwich ELISA and no HPT protein was detected. Conclusion CpTI protein content in transgenic rice seeds is lower than the detection limit, can not be quantified, and does not contain detectable levels of HPT protein.