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从大麦和小麦发育中的颖果分离的糊粉层原生质体中的瞬时基因表达。

Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat.

作者信息

Lee B, Murdoch K, Topping J, Kreis M, Jones M G

机构信息

AFRC Institute of Arable Crops Research, Harpenden, Herts, U.K.

出版信息

Plant Mol Biol. 1989 Jul;13(1):21-9. doi: 10.1007/BF00027332.

Abstract

Methods have been developed for the isolation of aleurone protoplasts from developing caryopses of Hordeum vulgare and Triticum aestivum in order to study transient expression of introduced genes. Chimaeric gene constructs were introduced into aleurone protoplasts by polyethylene glycol (PEG). Transient expression directed by the 35S promoter from cauliflower mosaic virus (CaMV) of the reporter gene encoding chloramphenicol acetyl transferase (CAT) was detected in aleurone protoplasts from developing barley and wheat grains. Using a similar construct, CAT activity increased when the alcohol dehydrogenase intron 1 fragment from maize was ligated between the 35S promoter and the CAT coding region. The demonstration of transient expression in protoplasts from developing aleurone layers indicates that they may be useful for investigating tissue and developmental control of genes coding for cereal seed proteins.

摘要

为了研究导入基因的瞬时表达,已开发出从大麦和小麦发育中的颖果中分离糊粉层原生质体的方法。通过聚乙二醇(PEG)将嵌合基因构建体导入糊粉层原生质体。在发育中的大麦和小麦籽粒的糊粉层原生质体中检测到由花椰菜花叶病毒(CaMV)的35S启动子指导的编码氯霉素乙酰转移酶(CAT)的报告基因的瞬时表达。使用类似的构建体,当将来自玉米的醇脱氢酶内含子1片段连接在35S启动子和CAT编码区之间时,CAT活性增加。在发育中的糊粉层的原生质体中瞬时表达的证明表明,它们可能有助于研究编码谷物种子蛋白的基因的组织和发育控制。

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