ISOLATION OF AN EFFICIENT ACTIN PROMOTER FOR USE IN RICE TRANSFORMATION
ISOLATION OF AN EFFICIENT ACTIN PROMOTER FOR USE IN RICE TRANSFORMATION
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DOI：
10.1105/tpc.2.2.163
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发表时间：
1990-02-01
期刊：
影响因子：
11.6
通讯作者：
WU, R
中科院分区：
文献类型：
--
作者：
MCELROY, D;ZHANG, WG;WU, R
我们对水稻肌动蛋白1基因（Act 1）的5“区进行了表征，表明它是一个有效的启动子，可调控外源基因在转基因水稻中的组成型表达。通过构建具有来自水稻Act 1基因的5“区域与编码细菌β-内酰胺酶的基因的编码序列融合的质粒，我们证明，Act 1翻译起始密码子上游的1.3个酶的区域含有高水平β-葡萄糖苷酸酶所必需的所有5“-调控元件。在转化的水稻原生质体的瞬时测定中检测葡糖醛酸酶（GUS）表达。水稻Act 1初级转录物具有一个非编码外显子，该外显子被一个5“内含子与第一编码外显子隔开。融合缺乏这个Act 1内含子表现出不可检测的GUS活性转化水稻原生质体的瞬时测定。Act 1 5“内含子的缺失分析表明，内含子介导的GUS表达刺激部分地与有效内含子剪接的体内需求相关。
We have characterized the 5'' region of the rice actin 1 gene (Act1) and show that it is an efficient promoter for regulating the constitutive expression of a foreign gene in transgenic rice. By constructing plasmids with 5'' regions from the rice Act1 gene fused to coding sequence of a gene encoding bacterial .beta.-glcuronidase, we demonstrate that a region 1.3 kilobases upstream of the Act1 translation initiation codon contains all of the 5''-regulatory elements necessary for high-level .beta.-glucuronidase (GUS) expression in transient assays of transformed rice protoplasts. The rice Act1 primary transcript has a noncoding exon separated by a 5'' intron from the first coding exon. Fusion that lack this Act1 intron showed no detectable GUS activity in transient assays of transformed rice protoplasts. Deletion analysis of the Act1 5'' intron suggests that the intron-mediated stimulation of GUS expression is associated, in part, with an in vivo requirement for efficient intron splicing.