含编码类铁氧还双亲性蛋白ap1基因的转基因水稻植株的获得(英文)

Production of Transgenic Rice Containing apl Gene Encoding a Ferredoxin-like Amphipathic Protein from Sweet Pepper

利用基因枪法将含有潮霉素抗性基因 (hpt)、gusA报告基因和ap1基因的 2个质粒 (pJIMB15和pBISAP1)共同转化由粳稻品种鄂宜 10 5号种子胚诱导的愈伤组织 (2～ 3周龄 ) .ap1基因编码一种双亲性的蛋白 .该蛋白能延缓因假单孢菌感染所引起的在非寄主植物中的过敏反应 .经过 2轮潮霉素 (30mg/L)筛选 ,抗性愈伤组织被转入含 30mg/L潮霉素的再生培养基中再生植株 .从轰击的 186块愈伤组织中共再生出 32株独立的转基因水稻植株 (转化率为 17.2 % ) .PCR/Southernblot分析显示 84%的转基因植株含有所有 3个基因 .

Two to three-week-old mature seed-derived rice callus (Oryza sativa L., japonica cv. Eyi 105) was co-transformed via particle bombardment with two plasmids, pJIMB15 and pBISAPl, containing the selectable marker hygromycin phosphotransferase gene (hpt), the reporter β-glucuronidase gene (gusA) and the ap1 gene. The ap1 gene encodes an amphipathic protein previously shown to delay the hypersensitive response induced in non-host plants by the pathogen Pseudomonas syringae pv. syringae(Pss). After two rounds of selection on hygromycin (30 mg/L)-containing medium, resistant callus was transferred to hygromycin (30 mg/L)-containing regeneration medium for plant regenaration. Thirty-two independent transgenic plants were regenerated from 186 bombarded callused, with a transformation frequency of 17.2%. Eighty-four percent of the transgenic plants carried all three transgenes as shown by PCR/Southern blot analysis.