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转基因技术的迅速发展有效地打破了物种间的遗传壁垒,从而加快了优良基因定向聚合的进程,然而遗传转化率低及转基因安全问题成为制约其进一步发展的限制因素。基于此,本研究以玉米自交系H99为材料,将Ac/Ds双元表达载体(包含抗虫Cry1 Ab/Ac基因和gfp报告基因等)通过农杆菌介导法转入由110粒玉米幼胚诱导出的愈伤组织中,获得28株抗性转基因植株。经PCR及RT-PCR的验证结果显示,其中8株为含有Cry1 Ab/Ac目的基因且该基因有效表达的阳性植株,转化效率达7%。通过农杆菌介导法所获得的玉米转基因植株为今后剔除抗生素筛选标记,从而获得安全的玉米抗虫新种质提供了遗传材料。
The rapid development of transgenic technology has effectively broken the genetic barriers among species, thus accelerating the progress of the excellent gene targeted polymerization. However, the low genetic transformation rate and the problem of transgenic safety have become the limiting factors restricting its further development. Based on this, the maize inbred line H99 was used as a material to transfer the Ac / Ds binary expression vector (including the insect-resistant Cry1 Ab / Ac gene and the gfp reporter gene) Embryo-induced callus, 28 resistant transgenic plants were obtained. The results of PCR and RT-PCR showed that 8 of them were positive plants containing Cry1 Ab / Ac gene and the expression of this gene was effective, and the transformation efficiency reached 7%. The maize transgenic plants obtained by the Agrobacterium-mediated method provide the genetic material for eliminating the antibiotic selection marker in the future, thereby obtaining the safe new corn germ-resistant germplasm.