Vianna G R, Aragão F J L, Rech E L
Embrapa Recursos Genéticos e Biotecnologia, Laboratório de Transferência e Expressão de Genes, Parque Estação Biológica, Brasília, DF, Brasil.
Genet Mol Res. 2011 Mar 1;10(1):382-90. doi: 10.4238/vol10-1gmr1058.
Currently, the market demands products committed to protecting human health and the environment, known as clean products. We developed a protocol using DNA fragments containing only the gene sequence of interest, to replace the circular vectors containing genes for antibiotic resistance and other undesirable sequences, for obtaining transgenic soybeans for microparticle bombardment. Vector pAC321 was digested with the restriction enzyme PvuII to produce the 6159 bp ahas fragment, which contains the mutated ahas gene from Arabidopsis thaliana (Brassicaceae), under the control of its own promoter and terminator. This gene confers resistance against imazapyr, a herbicidal molecule of the imidazolinone class, capable of systemically translocating and concentrating in the apical meristematic region of the plant, the same region used for the introduction of the transgenes. This fragment was used to generate 10 putative transgenic soybean lines.
目前，市场需要致力于保护人类健康和环境的产品，即清洁产品。我们开发了一种方案，使用仅包含目标基因序列的DNA片段，取代含有抗生素抗性基因和其他不良序列的环状载体，以获得用于微粒轰击的转基因大豆。用限制性内切酶PvuII消化载体pAC321，产生6159 bp的ahas片段，该片段包含来自拟南芥（十字花科）的突变ahas基因，受其自身启动子和终止子的控制。该基因赋予对咪草烟的抗性，咪草烟是咪唑啉酮类除草剂分子，能够在植物的顶端分生组织区域进行系统转运和积累，该区域也是用于导入转基因的区域。该片段用于产生10个推定的转基因大豆品系。
