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cry1Ab13基因是抗虫基因,编码的杀虫晶体蛋白对鳞翅目害虫具有一定的毒杀作用,构建了植物表达载体pCambia3300-35S-cry1Ab13,通过花粉管通道法将该载体转入大豆品种吉农28中,经PCR扩增检测得到15株T_1代阳性植株。Southern blotting分析显示有5株出现杂交信号,并以单拷贝形式整合到大豆基因组中。荧光定量PCR测定结果表明:cry1Ab13基因在转化植株的叶、茎秆中均有表达,每株表达量各不相同,在叶片部位表达量相对较高,最高为6.7,最低为2.5;在茎部表达量最高为0.76,最低为0.31。对T_1代阳性植株籽粒,采用圆盘分隔法接入大豆食心虫幼虫,进行初步抗虫试验,结果显示转化植株抗虫效果明显,但后代稳定性还需进一步研究。
The cry1Ab13 gene is an insect-resistant gene, and the encoded insecticidal crystal protein has some toxic effects on Lepidoptera pests. The plant expression vector pCambia3300-35S-cry1Ab13 was constructed and transformed into the soybean variety Kyongon through pollen tube pathway 28, 15 T_1 generation positive plants were obtained by PCR amplification. Southern blotting analysis showed that 5 out of hybridization signals, and a single copy of the integration into the soybean genome. The result of real-time quantitative PCR showed that cry1Ab13 gene was expressed in the leaves and stems of transformed plants, and the expression level of each strain was different. The expression level of cry1Ab13 gene in leaves was relatively high, the highest was 6.7 and the lowest was 2.5. The highest expression level was 0.76 and the lowest level was 0.31. The T 1 generation positive plants were infected with soybean borer larvae by disk separation method, and the preliminary pest resistance test showed that the transgenic plants had obvious anti-insect effect, but the stability of progeny should be further studied.