电穿孔介导的原生质体直接基因转移法获得的白茅转基因植株。

Transgenic plants of Agrostis alba obtained by electroporation-mediated direct gene transfer into protoplasts.

机构信息

Faculty of Horticulture, Chiba University, 6-2-1 Kashiwa-no-ha, Kashiwa-city, 277, Chiba, Japan.

出版信息

Plant Cell Rep. 1994 Feb;13(5):243-6. doi: 10.1007/BF00233312.

PMID:24193757

Abstract

A system for genetic transformation of Agrostis alba plants by electroporation-mediated DNA transfer to protoplasts is described. The npt II gene was used as a selectable marker. Selection with 20 mg/1 G418 (geneticin) yielded a total of over 50 resistant cell colonies from three independent experiments. Overall frequency of resistant colony formation was 1-3 × 10(-6) based on the number of protoplasts plated and 1-2 × 10(-5) based on the number of cell colonies recovered. Subsequent subcultures led to the development of plants with an apparently normal morphology. DNA analysis (PCR and Southern hybridization) and enzymatic analysis showed that the G418 resistant plants carried the transgene and expressed it. This is the first successful genetic transformation of an economically important temperate grass, Agrostis.

摘要

描述了用电穿孔介导的 DNA 转移原生质体将遗传转化系统引入白茅草植物的方法。npt II 基因被用作选择标记。用 20mg/1 G418（遗传霉素）选择，从三个独立的实验中总共获得了超过 50 个抗性细胞集落。根据平板原生质体的数量，抗性集落形成的总频率为 1-3×10(-6)，根据回收的细胞集落的数量，频率为 1-2×10(-5)。随后的继代培养导致具有明显正常形态的植物的发育。DNA 分析（PCR 和 Southern 杂交）和酶分析表明，G418 抗性植物携带转基因并表达它。这是首例成功的经济上重要的温带草，白茅草的遗传转化。