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首页> 外文期刊>Journal of Turfgrass Management >Transient Expression of the Beta-Glucuronidase Gene Driven by Different 5' Regulatory Sequences in Tall Fescue Protoplasts
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Transient Expression of the Beta-Glucuronidase Gene Driven by Different 5' Regulatory Sequences in Tall Fescue Protoplasts

机译：高羊茅原生质体中不同的5'调控序列驱动的β-葡萄糖醛酸酶基因的瞬时表达。

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摘要

To identify strongly active 5' regulatory sequences for high levels of transgene expression in tall fescue (Festuca arundina-cea Schreb.), we have tested the cauliflower mosaic virus (CaMV) 35S promoter and four different promoter/intron constructs: the CaMV 35S promoter plus the maize alcohol dehydrogenase 1 (adhl) gene intron 1, the CaMV 35S promoter plus the maize adhl intron 6, the rice actin 1 promoter plus its first, intron, and the maize adhl promoter plus its first intron. These 5' regulatorysequences were connected to the beta-glucuronidase (GUS) gene for a convenient assay. After the plasmids containing these constructs were introduced into tall fescue protoplasts by electroporation, transient GUS expression was measured to determine their activities. Among the various constructs tested, the rice actin 1 promoter and its first intron showed the highest GUS expression, followed by the CaMV 35S promoter plus maize adhl intron 6. The maize adhl promoter and its first intron showed intermediate GUS expression whereas the CaMV 35S promoter alone or plus the maize adhl intron 1 exhibited very weak GUS expression in tall fescue.
机译：为了确定高活性羊茅（Festuca arundina-cea Schreb。）中高水平转基因表达的强活性5'调控序列，我们测试了花椰菜花叶病毒（CaMV）35S启动子和四种不同的启动子/内含子构建体：CaMV 35S启动子加上玉米醇脱氢酶1（adhl）基因内含子1，CaMV 35S启动子加上玉米adhl内含子6，水稻肌动蛋白1启动子加上它的第一个内含子，以及玉米adhl启动子加上它的第一个内含子。将这些5'调节序列连接至β-葡糖醛酸糖苷酶（GUS）基因以方便测定。通过电穿孔将含有这些构建体的质粒引入高羊茅原生质体后，测量瞬时GUS表达以确定其活性。在测试的各种构建体中，水稻肌动蛋白1启动子及其第一个内含子显示出最高的GUS表达，随后是CaMV 35S启动子加上玉米adhl内含子6。玉米adhl启动子及其第一个内含子显示出中等的GUS表达，而CaMV 35S启动子单独或加玉米的adhl内含子1在高羊茅中表现出非常弱的GUS表达。

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