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[利用嵌合GFP-TUA6基因对亚麻(Linum usitatissimum L.)进行遗传转化以可视化微管]

[Genetic transformation of flax (Linum usitatissimum L.) with chimeric GFP-TUA6 gene for visualisation of microtubules].

作者信息

Shisha E N, Korkhovoĭ V I, Baer G Ia, Guzenko E V, Lemesh V A, Kartel' N A, Emets A I, Blium Ia B

出版信息

Tsitol Genet. 2013 Mar-Apr;47(2):3-11.

Abstract

The data of Agrobacterium-mediated transformation of some Linum usitatissimum cultivars zoned on the territories of Belarus and Ukraine with the plasmid carrying chimeric GFP-TUA6 gene and nptII gene as selectable marker conferring resistance to kanamycin are presented in this study. Transformation was affected by a number of factors including optical density (OD600), time of inoculation of explants with Agrobacterium and co-culture conditions. Transgenic nature of obtained lines was confirmed by PCR analysis. Expression of GFP-TUA6 gene was detected with confocal laser scanning microscopy. The obtained transgenic lines can be used for further functional studies the role of microtubules in the processes of building the flax fibres and resistance to wind.

摘要

本研究展示了利用携带嵌合绿色荧光蛋白-微管蛋白α6(GFP-TUA6)基因的质粒以及赋予卡那霉素抗性的nptII基因作为选择标记,对白俄罗斯和乌克兰境内种植的一些亚麻品种进行农杆菌介导转化的数据。转化受到多种因素影响,包括光密度(OD600)、外植体与农杆菌接种时间以及共培养条件。通过聚合酶链反应(PCR)分析证实了所得株系的转基因性质。利用共聚焦激光扫描显微镜检测了GFP-TUA6基因的表达。所获得的转基因株系可用于进一步开展功能研究,以探究微管在亚麻纤维形成过程中的作用以及对风的抗性。

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