RAPID TRANSIENT INDUCTION OF PHENYLALANINE AMMONIA-LYASE MESSENGER-RNA IN ELICITOR-TREATED BEAN CELLS
RAPID TRANSIENT INDUCTION OF PHENYLALANINE AMMONIA-LYASE MESSENGER-RNA IN ELICITOR-TREATED BEAN CELLS
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DOI：
10.1073/pnas.82.20.6731
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
LAMB, CJ
中科院分区：
文献类型：
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作者：
EDWARDS, K;CRAMER, CL;LAMB, CJ
与存在于激发子处理的菜豆（Phaseolus vulgaris L.）细胞中的聚（A）+ RNA的大小选择部分互补的DNA。插入到pAT 153中，并用于转化大肠杆菌菌株C600。通过杂交选择翻译和交叉杂交鉴定了5个克隆，其含有与编码苯丙氨酸解氨酶（EC 4.3.1.5）的mRNA互补的序列，所述苯丙氨酸解氨酶催化苯丙素类生物合成的第一反应。最长的插入片段含有一个1520个碱基对的开放阅读框和223个碱基对的3“非翻译序列。RNA印迹杂交表明，诱导子引起苯丙氨酸解氨酶mRNA的快速，显着，但短暂的增加，这是密切相关的在体外和体内酶合成的可翻译的mRNA活性的变化。新合成的mRNA纯化的有机汞亲和层析后，在体内脉冲标记4-硫尿苷的印迹杂交表明，激发子引起的苯丙氨酸解氨酶mRNA合成的快速刺激作为一个早期事件的防御反应，导致苯丙烷衍生的植物抗毒素的积累。
DNAs complementary to a size-selected fraction of poly(A)+ RNA present in elicitor-treated cells of bean (Phaseolus vulgaris L.) were inserted into pAT153 and used to transform Escherichia coli strain C600. Five clones were identified by hybrid-selected translation and cross-hybridization that contained sequences complementary to mRNA encoding phenylalanine ammonia-lyase (EC 4.3.1.5), which catalyzes the first reaction of phenylpropanoid biosynthesis. The longest insert contained a single open reading frame of 1520 base pairs together with 223 base pairs of 3'' untranslated sequence. RNA blot hybridization showed that elicitor caused a rapid, marked but transient increase in phenylalanine ammonia-lyase mRNA that was closely correlated with changes in translatable mRNA activity in vitro and enzyme synthesis in vivo. Blot hybridization of newly synthesized mRNA purified by organomercurial affinity chromatography following in vivo pulse-labeling with 4-thiouridine indicates that elicitor caused a rapid stimulation of phenylalanine ammonia-lyase mRNA synthesis as an early event in the defense response leading to accumulation of phenylpropanoid-derived phytoalexins.