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Bt cry1Ah基因是从国内苏云金芽胞杆菌分离株BT8中鉴定克隆的新型杀虫蛋白基因.本研究构建了两个含有Bt cry1Ah基因的植物表达载体,在其中一个植物表达载体(pUUOAH)的玉米Ubiquitin启动子与cry1Ah基因之间插入了玉米泛素蛋白基因ubiqutin1的第一个内含子(ubi1).利用基因枪法将其导入玉米(ZeamaysL.)胚性愈伤组织中,得到了可育的再生植株.PCR及Southern blot检测结果表明,cry1Ah基因已整合入玉米基因组中,并稳定遗传至下一代.对T1及T2代转基因植株进行ELISA检测,发现pUUOAH载体转基因植株Cry1Ah蛋白的平均表达量提高了20%.对T2代转基因植株进行了田间抗虫性鉴定,发现转cry1Ah基因玉米对玉米螟有很高的抗性,并且pUUOAH载体转基因植株的抗虫性好于pUOAH(无内含子)载体的转基因植株.以上结果表明,玉米ubi1内含子可以有效增强cry1Ah在转基因玉米中的表达。
The Bt cry1Ah gene is a novel insecticidal protein gene identified from the domestic Bacillus thuringiensis isolates BT8.In this study, two plant expression vectors containing the Bt cry1Ah gene were constructed, and in which Ubiquitin, a maize plant expression vector (pUUOAH) The first intron (ubi1) of ubiqutin1 was inserted between cry1Ah gene and cry1Ah gene and introduced into embryogenic callus of maize (Zea mays L.) by particle bombardment to obtain fertile regenerated plants The result of PCR and Southern blot showed that the cry1Ah gene was integrated into the genome of maize and was inherited stably to the next generation.The ELISA of T1 and T2 transgenic plants showed that the average expression level of Cry1Ah protein in transgenic plants of pUUOAH vector increased by 20 % .Two generations of transgenic plants were identified for their resistance to insecticides. The results showed that transgenic corn with cry1Ah gene was highly resistant to Ostrinia furnacalis and the insect resistance of transgenic plants with pUUOAH vector was better than that of pUOAH (without intron) Transgenic plants.The above results show that the maize ubi1 intron can effectively enhance the cry1Ah expression in transgenic maize.