Nagashima Yukihiro, Koiwa Hisashi
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station 77843-2133, TX, USA.
Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station 77843-2133, TX, USA; Molecular and Environmental Plant Science Program, Texas A&M University, College Station 77843-2133, TX, USA.
Anal Biochem. 2017 May 15;525:44-45. doi: 10.1016/j.ab.2017.02.017. Epub 2017 Feb 27.
Kanamycin resistance is the most frequently used antibiotic-resistance marker for Arabidopsis transformations, however, this method frequently causes escape of untransformed plants, particularly at the high seedling density during the selection. Here we developed a robust high-density selection method using top agar for Arabidopsis thaliana. Top agar effectively suppressed growth of untransformed wild-type plants on selection media at high density. Survival of the transformed plants during the selection were confirmed by production of green true leaves and expression of a firefly luciferase reporter gene. Top agar method allowed selection using a large amount of seeds in Arabidopsis transformation.
卡那霉素抗性是拟南芥转化中最常用的抗生素抗性标记,但这种方法经常导致未转化植株逃逸,尤其是在选择过程中幼苗密度较高时。在此,我们开发了一种使用顶层琼脂的稳健的高密度选择方法用于拟南芥。顶层琼脂有效地抑制了未转化野生型植株在高密度选择培养基上的生长。通过绿色真叶的产生和萤火虫荧光素酶报告基因的表达证实了转化植株在选择过程中的存活。顶层琼脂法允许在拟南芥转化中使用大量种子进行选择。