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高粱是全球仅次于小麦、水稻、玉米、大豆和马铃薯等的重要作物之一。以高粱幼穗愈伤组织为转化受体,通过农杆菌介导法和含有抗潮霉素和gus基因的双元载体将杀虫晶体蛋白基因cry1Ab导入高粱品种115、ICS21B和5-27,经Hyg筛选共获得21个独立的转基因株系,52株转基因植株,平均转化率为1.9%。经PCR、Southern杂交和RT-PCR分析表明cry1Ab基因已整合入高粱基因组中并得到正确转录。Bt蛋白Westernblotting分析和ELISA定量测定显示,cry1Ab基因在转基因高粱植株中表达,但不同转基因植株表达量有差异。饲虫试验表明,转基因高粱对大螟(Sesamiainferens)具有一定抗性。
Sorghum is one of the most important crops in the world after wheat, rice, corn, soybean and potato. The sorghum young calli were used as the transformation recipients and the insecticidal crystal protein gene cry1Ab was introduced into sorghum variety 115, ICS21B and 5-27 via Agrobacterium-mediated method and binary vector containing hygromycin resistance and gus gene. Hyg screened a total of 21 independent transgenic lines, 52 transgenic plants, the average conversion rate of 1.9%. PCR, Southern blot and RT-PCR analysis showed that the cry1Ab gene has been integrated into the sorghum genome and correctly transcribed. Western blot analysis of Bt protein and ELISA assay showed that cry1Ab gene was expressed in transgenic sorghum plants, but the expression levels of different transgenic plants were different. Insecticide tests showed that transgenic sorghum has some resistance to Sesamia inferens.