Wang G, Zhang H, Ding Q, Xie Y, Dai J
College of Biology, Beijing Agricultural University, China.
Chin J Biotechnol. 1996;12(1):53-60.
Maize transformation has been carried out through microprojectile bombardment, ultrasonication in a DNA buffer, and ovary-injection with a self-made microinjector. The plasmid pB48.415, which carries a 3'-truncated Bt-toxin protein gene and a hygromycin phosphotransferase (hpt) gene, was used in the transformation. Transgenic maize plants were obtained from immature embryos and embryogenic calli bombarded with a particle gun, embryogenic calli ultrasonicated under different conditions of ovaries injected 10-20 hours after pollination. The results of Dot blotting and Southern blotting analyses proved the integration of the Bt gene into maize genome.
玉米转化已通过微粒轰击、在DNA缓冲液中进行超声处理以及使用自制微注射器进行子房注射来实现。携带3'端截短的Bt毒素蛋白基因和潮霉素磷酸转移酶(hpt)基因的质粒pB48.415被用于转化。通过用粒子枪轰击未成熟胚和胚性愈伤组织、在授粉后10 - 20小时对不同条件下的子房进行超声处理后的胚性愈伤组织,获得了转基因玉米植株。斑点杂交和Southern杂交分析结果证明了Bt基因已整合到玉米基因组中。