公开(公告)号：US08030543B2

公开(公告)日：2011-10-04

申请号：US12087208

申请日：2006-01-12

申请人： Villoo Morawala Patell ,  Rajyashri Ramakrishna Karur

发明人： Villoo Morawala Patell ,  Rajyashri Ramakrishna Karur

IPC分类号： C12N15/82 ,  C12N15/63 ,  C12N15/00

CPC分类号： C12N15/821 ,  C12N9/92

摘要： The invention describes a highly improved, reproducible and a consistent method of transformation and regeneration that results in obtaining 12-15% transgenic plants. The present invention relates to a method of selecting genetically transformed Sunflower explants based on their ability to utilize Xylose as a sole carbohydrate source. Further disclosed is the nucleic acid sequence of the Xylose Isomerase gene, vector construction for incorporation of the selection marker gene and the process of Agrobacterium Mediated Transformation of target host plant with the vector comprising the gene encoding the enzyme Xylose isomerase under the functional combination of the heterologous regulatory sequences. Also disclosed is the method of selecting the putative transformants post transformation with the said vector that possess a metabolic advantage of utilizing Xylose as a sole carbon source. Increased efficiency of regeneration, better growth and survival has been observed on subjection to the described method of positive selection. The subject invention alleviates the disadvantages of negative selection methods such as the undesired elimination of the transformed cells and the potential environmental harm caused due to the dispersal of the antibiotic and the herbicide resistant genes.

摘要翻译： 本发明描述了高度改进的，可重复的和一致的转化和再生方法，其导致获得12-15％的转基因植物。 本发明涉及一种基于其利用木糖作为唯一碳水化合物来源的能力来选择遗传转化的向日葵外植体的方法。 进一步公开的是，在包含编码酶木糖异构酶的基因的载体的功能组合下，将木糖异构酶基因的核酸序列，掺入选择标记基因的载体构建和农杆菌介导的靶宿主植物转化过程 异源调控序列。 还公开了使用具有利用木糖作为唯一碳源的代谢优势的所述载体选择转化后的推定转化体的方法。 对于所选择的阳性选择方法，观察到提高再生效率，更好的生长和生存。 本发明减轻了阴性选择方法的缺点，例如不希望地消除转化细胞，以及由于抗生素和除草剂抗性基因的分散引起的潜在的环境危害。