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首页> 外文期刊>Acta Horticulturae >Agrobacterfum-medlated genetic transformation of 146-2 Russian apricot and plum rootstock with the gfp gene
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Agrobacterfum-medlated genetic transformation of 146-2 Russian apricot and plum rootstock with the gfp gene

机译：Agrobacterfum-Medlated遗传转化为146-2俄罗斯杏和李子砧木与GFP基因

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摘要

A protocol for Agrobacterium-mediated genetic transformation was developed for 146-2 Russian apricot rootstock. Whole leaves from in vitro cultured shoots were used as explants for transformation by A tumefaciens CBE21 strain containing binary vectorpBINmGFPSER. It has the nptll gene coding for neomycin phosphotransferase II as a plant-selectable marker under the control of the NOS (nopaline synthase) promoter and gfp reporter gene coding the green fluorescent protein under the control of the Cauliflower mosaic virus (CaMV) 35S promoter. Integration of the nptll and gfp into transgenes was confirmed by PCR. The expression of the green fluorescent protein was observed by fluorescence microscopy. Transformation efficiency based on PCR analysis of kanamycin resistant independent lines was 0.41-0.83%. All transgenic lines showed kanamycin resistance at concentration 40 mg L1. They were rooted and acclimatized to greenhouse conditions.
机译：制定了146-2俄罗斯砧木的农杆菌介导的遗传转化的方案。 来自体外培养芽的整个叶子用作通过含有二元瓣糖浆的肿瘤CBE21菌株转化的外植体。 它具有在NoS（Nopaline合成酶）启动子（Cauliflower Mosaic病毒（CAMV）35s启动子的控制下编码绿色荧光蛋白的NOS（Nopaline）启动子和GFP报告基因的植物可选标记的NPTLL基因编码。 通过PCR证实了NPTLL和GFP将NPTL和GFP的整合。 通过荧光显微镜观察绿色荧光蛋白的表达。 基于PCR分析的Kanamycin抗性独立线的转化效率为0.41-0.83％。 所有转基因系列浓度为40mg L1时显示出Kanamycin抗性。 他们根植和适应温室条件。

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  • 来源
    《Acta Horticulturae》 |2020年第1282期|共6页
  • 作者

    L.Y. Mourenets; A.S. Pushin; S.V. Dolgov;

  • 作者单位

    Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences Pushchino Russia;

    Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences Pushchino Russia;

    Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences Pushchino Russia;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 园艺;
  • 关键词

    leaf explants; CBE21; A. tumefaciens; nptll; GFP.;

    机译：叶子外植物;cbe21;a。 Tumefaciens;NPTLL;GFP。;

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