转基因抗虫油菜中Bt杀虫蛋白基因稳定遗传和高效表达及抗虫性研究

Stable Heredity and Efficient Expression of Bt Insecticidal Protein Gene in the Transgenic Rapeseed and Its Insect-resistant Activity

通过对转 Bt杀虫蛋白基因油菜植株的后代进行卡那霉素抗性分析和 PCR技术检测 ,结果表明 :Bt杀虫蛋白基因是以单拷贝、杂合地整合到转基因植株 (T0 1、T0 2、T0 3、T0 5、T0 6、T0 7、T0 8)的基因组中 ,并稳定地遗传。EL ISA检测表明 :在第 3～第 9叶期 Bt杀虫蛋白基因在转基因油菜植株中高效地表达 ,Bt杀虫蛋白的表达量为 170～ 2 6 0 ng/ 2 5mg鲜叶 ,占植物可溶性蛋白的 0 .0 6 7%～ 0 .10 5 % ,其抗虫效果高达 72 .0 %～ 94 .4 %。但从第 11叶期后 Bt杀虫蛋白表达量显著地降低 ,只有 7～ 5 0 ng/ 2 5 mg鲜叶 ,占植物可溶性蛋白的 0 .0 0 3%～ 0 .0 2 %。

The progeny of transgenic rapeseed plants with Bt insecticidal protein gene was detected by kana mycin sifting and PCR. Results showed that single copy of Bt insecticidal protein gene has been integrated into the genome of T 0 transgenic plants and stably inherited.By ELISA Bt insecticidal protein gene can efficiently express in the transgenic rapeseed plants with the 3th^9th leaf stage, and the expressing quantity is 170~260ng/25mg fresh leaf.It amounts to 0.067%~0.105% of plant soluble protein,and the insecticidal activity is 72.0%~99.4%.But expressing quantity of Bt insecticidal protein apparently decreased after the 11th leaf stage and down to 7~50ng/25mg fresh leaf,corresponding to 0.003%~0.02% of plant soluble protein.