Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana
Molecular cloning and expression analysis of the mevalonate kinase gene from Arabidopsis thaliana
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DOI：
10.1023/a:1006325630792
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发表时间：
2000-01-01
影响因子：
5.1
通讯作者：
Ferrer, A
中科院分区：
文献类型：
--
作者：
Lluch, MA;Masferrer, A;Ferrer, A
甲羟戊酸激酶（MVK）是一类催化甲羟戊酸磷酸化生成5-磷酸甲羟戊酸的酶，被认为是类异戊二烯生物合成途径的潜在调控酶。已克隆并鉴定了与先前分离的MVK cDNA相对应的拟南芥MVK基因。RNA酶保护分析表明，MVK基因的表达产生三个mRNA群体，其5'端定位于MVK ATG起始密码子上游203、254和355 nt。北方杂交分析表明，MVK mRNA在根和花序中优先积累。组织化学分析，转基因A。含有MVK基因5 ′区的1.8kb片段与β-葡萄糖醛酸酶（GUS）报告基因的翻译融合体的拟南芥植物表明，MVK 5 ′侧翼区指导GUS基因在整个发育过程中的广泛表达，尽管在根（分生区）和花（萼片、花瓣、花药、花柱和柱头乳突）中检测到最高水平的GUS活性。MVK基因的表达模式表明，编码的MVK的作用是产生甲羟戊酸-5-磷酸的一般池，用于合成涉及基本和专门植物细胞功能的不同类异戊二烯。在转染的A.拟南芥原生质体表明MVK 5 ′-侧翼区-295和-194位之间的调节元件对于高水平MVK基因表达是至关重要的。
Mevalonate kinase (MVK), the enzyme that catalyzes the phosphorylation of mevalonate to produce mevalonate 5-phosphate, is considered as a potential regulatory enzyme of the isoprenoid biosynthetic pathway. The Arabidopsis thaliana MVK gene corresponding to the MVK cDNA previously isolated has been cloned and characterized. RNAse protection analysis indicated that the expression of the MVK gene generates three mRNA populations with 5' ends mapping 203, 254 and 355 nt upstream of the MVK ATG start codon. Northern blot analysis showed that the MVK mRNA accumulates preferentially in roots and inflorescences. Histochemical analysis, with transgenic A. thaliana plants containing a translational fusion of a 1.8 kb fragment of the 5' region of the MVK gene to the beta-glucuronidase (GUS) reporter gene, indicated that the MVK 5'-flanking region directs widespread expression of the GUS gene throughout development, although the highest levels of GUS activity are detected in roots (meristematic region) and flowers (sepals, petals, anthers, style and stigmatic papillae). The expression pattern of the MVK gene suggests that the role of the encoded MVK is the production of a general pool of mevalonate-5-phosphate for the synthesis of different classes of isoprenoids involved in both basic and specialized plant cell functions. Functional promoter deletion analysis in transfected A. thaliana protoplasts indicated that regulatory elements between positions -295 and -194 of the MVK 5'-flanking region are crucial for high-level MVK gene expression.