Generation and analysis of soybean plastid transformants expressing Bacillus thuringiensis Cry1Ab protoxin
Generation and analysis of soybean plastid transformants expressing Bacillus thuringiensis Cry1Ab protoxin
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DOI：
10.1007/s11103-005-7405-3
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发表时间：
2005-07-01
影响因子：
5.1
通讯作者：
Dubald, M
中科院分区：
文献类型：
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作者：
Dufourmantel, N;Tissot, G;Dubald, M
我们描述了一代可育和同质大豆质体转化体，表达苏云金芽孢杆菌杀虫原毒素Cry1Ab。将转基因靶向大豆质体的基因间区域，在rps1217和trnV基因之间，并使用编码壮观霉素抗性的aadA基因进行选择。分子分析证实了cry1Ab和aadA表达盒在大豆质体中的预期位置处的整合，以及转基因到下一代的传递。Western blot分析表明Cry1Ab原毒素在叶、茎和种子中高表达，但在根中不表达。它的表达赋予所产生的转基因大豆强的杀虫活性，如以绒豆毛虫（黎豆夜蛾（Anticarsia gemmatalis））为例。
We describe the generation of fertile and homoplasmic soybean plastid transformants, expressing the Bacillus thuringiensis insecticidal protoxin Cry1Ab. Transgenes were targeted in the intergenic region of Glycine max plastome, between the rps12\7 and trnV genes and selection was carried out using the aadA gene encoding spectinomycin resistance. Molecular analysis confirmed the integration of the cry1Ab and aadA expression cassettes at the expected location in the soybean plastome, and the transmission of the transgenes to the next generation. Western blot analyses showed that the Cry1Ab protoxin is highly expressed in leaves, stems and seeds, but not in roots. Its expression confers strong insecticidal activity to the generated transgenic soybean, as exemplified with velvetbean caterpillar (Anticarsia gemmatalis).