Imperial Chemical Industries, Corporate Biosciences Laboratory, The Health, Runcorn, Cheshire WA47 4QE, United Kingdom.
Proc Natl Acad Sci U S A. 1985 Oct;82(20):6731-5. doi: 10.1073/pnas.82.20.6731.
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 in the defense response leading to accumulation of phenylpropanoid-derived phytoalexins.
与激发子处理的菜豆（Phaseolus vulgaris L.）细胞中多聚（A）（+）RNA 的一个大小选择部分互补的 DNA 被插入 pAT153 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，它们包含与编码苯丙氨酸氨裂解酶（EC 4.3.1.5）的 mRNA 互补的序列，该酶催化苯丙烷生物合成的第一个反应。最长的插入物包含一个 1520 个碱基对的单一开放阅读框，以及 223 个碱基对的 3'非翻译序列。RNA 印迹杂交显示，激发子引起苯丙氨酸氨裂解酶 mRNA 的快速、显著但短暂的增加，这与体外可翻译 mRNA 活性和体内酶合成的变化密切相关。用 4-硫代尿嘧啶对体内脉冲标记后通过有机汞亲和层析纯化的新合成的 mRNA 的印迹杂交表明，激发子引起苯丙氨酸氨裂解酶 mRNA 合成的快速刺激，作为防御反应的早期事件，导致苯丙烷衍生的植物抗毒素的积累。
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