文献类型： 外文期刊

作者： Chen, SB ¹ ; Li, XG ² ; Liu, X ² ; Xu, HL ² ; Meng, K ² ; Xiao, GF ² ; Wei, XL ³ ; Wang, F; Zhu, Z;

作者机构： 1.Chinese Acad Sci, Inst Genet & Dev Biol, Beijing 100101, Peoples R China

2.Chinese Acad Sci, Inst Genet & Dev Biol, Beijing 100101, Peoples R China; Fujian Acad Agr Sci, Fujian Prov Key Lab Agr Genet Engn, Fuzhou 350003, Peoples R China

3.Chinese Acad Sci, Inst Genet & Dev Biol, Beijing 100101, Peoples R China; Fujian Acad Agr Sci, Fujian Prov Key Lab Agr Genet Engn, Fuzh

关键词： co-transformation;marker-free;double T-DNA vector;green fluorescent protein;Nicotiana tabacum

期刊名称：PLANT CELL REPORTS （ 影响因子：4.57； 五年影响因子：4.463 ）

ISSN： 0721-7714

年卷期： 2005 年 23 卷 9 期

页码：

收录情况： SCI

摘要： We investigated the potential of a novel double T-DNA vector for generating marker-free transgenic plants. Co-transformation methods using a double T-DNA vector or using mixture of two Agrobacterium tumefaciens strains were compared, and showed that the double T-DNA vector method could produce marker-free transgenic tobacco (Nicotiana tabacum L.) plants more efficiently. A dual marker double T-DNA vector was then constructed by assembling the green fluorescent protein (GFP) gene mgfp5 and the neomycin phosphotransferase gene nptII into the same T-DNA. The frequency of co-transformants produced by this vector was 56.3%. Co-expression of mgfp5 and nptII was found in 28 out of 29 T-1 lines, and segregation of the reporter beta-glucuronidase gene, gusA, from mgfp5 to nptII was found in 12 out of 29 T-1 lines. Therefore, GFP could be used as a vital marker to improve the transformation efficiency and to easily monitor the segregation of marker genes, thus facilitating screening of marker-free progeny.