通过对表达绿色荧光蛋白的细胞进行视觉筛选获得转基因燕麦植株。

Transgenic oat plants via visual selection of cells expressing green fluorescent protein.

作者信息

Kaeppler H F, Menon G K, Skadsen R W, Nuutila A M, Carlson A R

机构信息

Department of Agronomy, University of Wisconsin, 1575 Linden Drive, Madison, WI 53706, USA e-mail:

Novartis Seeds, Inc., 3054 Cornwallis Road, Research Triangle Park, NC 27709, USA, , , , , , WF.

出版信息

Plant Cell Rep. 2000 Jun;19(7):661-666. doi: 10.1007/s002999900167.

DOI:10.1007/s002999900167

PMID:30754802

Abstract

New selectable markers and selection systems are needed to increase the efficiency and flexibility of plant transformation. The objective of this research was to determine if the green fluorescent protein (gfp) gene could be utilized as a visual selectable marker for transformation of oat (Avena sativa L.). A modified gfp gene was delivered into oat cells by microprojectile bombardment. Cell clusters expressing gfp were visually identified using fluorescence microscopy and physically isolated at each subculture. Eleven independent transgenic cell lines were obtained, and fertile plants regenerated from all lines. Transgene integration and expression were confirmed in transgenic plants and progeny. Transgene expression segregated in a 3 : 1 ratio in progeny of the majority of the transgenic lines.

摘要

需要新的选择标记和选择系统来提高植物转化的效率和灵活性。本研究的目的是确定绿色荧光蛋白（gfp）基因是否可用作燕麦（Avena sativa L.）转化的视觉选择标记。通过微粒轰击将一个修饰的gfp基因导入燕麦细胞。使用荧光显微镜目视鉴定表达gfp的细胞团，并在每次继代培养时进行物理分离。获得了11个独立的转基因细胞系，所有系均再生出可育植株。在转基因植物及其后代中证实了转基因的整合和表达。大多数转基因系的后代中，转基因表达以3∶1的比例分离。