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用于转基因植物中转化、异源基因表达及转座子切除检测的有效载体。

Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants.

作者信息

Jones J D, Shlumukov L, Carland F, English J, Scofield S R, Bishop G J, Harrison K

机构信息

John Innes Centre for Plant Science Research, Norwich Research Park, Colney, UK.

出版信息

Transgenic Res. 1992 Nov;1(6):285-97. doi: 10.1007/BF02525170.

Abstract

Progress in plant molecular biology has depended heavily on the availability of effective vectors for plant cell transformation and heterologous expression. In this paper we describe the structures of a wide array of plasmids which have proved extremely effective in (a) plant transformation, (b) expression of heterologous genes and (c) assaying the activity of transposons in transgenic plants. Constructs that confer resistance to kanamycin, hygromycin, streptomycin, spectinomycin and phosphinotricin, or that confer beta-glucuronidase (GUS) gene expression are presented. Binary vector constructs that carry polylinkers of the pUC and Bluescript types are also described. Plasmids that permit the expression of any heterologous reading frame from either nopaline synthase (nos) or octopine synthase (ocs) promoters, as well as the cauliflower mosaic virus 35S promoter, using either the nopaline synthase or octopine synthase 3' polyadenylation sequences, are presented. These constructs permit a choice of orientation of the resulting transgene of interest, relative to the orientation of the selection marker gene. Most of the plasmids described here are publicly available.

摘要

植物分子生物学的进展在很大程度上依赖于用于植物细胞转化和异源表达的有效载体。在本文中,我们描述了一系列质粒的结构,这些质粒已被证明在以下方面极为有效:(a)植物转化,(b)异源基因表达,以及(c)在转基因植物中检测转座子的活性。展示了赋予对卡那霉素、潮霉素、链霉素、壮观霉素和草丁膦抗性的构建体,或赋予β-葡萄糖醛酸酶(GUS)基因表达的构建体。还描述了携带pUC和蓝思载体类型多克隆位点的双元载体构建体。展示了使用胭脂碱合成酶或章鱼碱合成酶3'聚腺苷酸化序列,允许从胭脂碱合成酶(nos)或章鱼碱合成酶(ocs)启动子以及花椰菜花叶病毒35S启动子表达任何异源阅读框的质粒。这些构建体允许所产生的目的转基因相对于选择标记基因的方向进行选择。这里描述的大多数质粒都是公开可用的。

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