Sokhansandzh A, Neumyvakin L V, Moseĭko N A, Piruzian E S
Genetika. 1997 Jul;33(7):906-13.
Accumulation of free proline (Pro) by bacteria and plants serves as a mechanism protecting these organisms from abiotic stress. We introduced two E. coli genes, proBosm, encoding a mutant variant of the first enzyme of the Pro biosynthetic pathway, gamma-glutamyl kinase, and proA, encoding the second enzyme of the Pro pathway, glutamyl-gamma-semialdehyde dehydrogenase, into tobacco plants. The spontaneous proBosm mutation was selected for Pro overproduction and mapped to the N-terminus of a polypeptide chain; it was found to result from one amino-acid substitution. Two E. coli genes were simultaneously introduced into tobacco plants, each under the control of a strong constitutive CaMV 35S promoter, which contained a duplicated sequence of enhancer, or the root-specific Pmas promoter sequence. Transgenic plants were characterized by Pro overproduction, increased resistance to L-azetidine-2-carboxylic acid (a toxic analog of Pro), and tolerance to salt stress.
细菌和植物积累游离脯氨酸(Pro)是这些生物体抵御非生物胁迫的一种机制。我们将两个大肠杆菌基因导入烟草植株,其中proBosm编码脯氨酸生物合成途径中第一种酶γ-谷氨酰激酶的突变变体,proA编码脯氨酸途径中的第二种酶谷氨酰-γ-半醛脱氢酶。为了实现脯氨酸的过量生产,我们选择了proBosm的自发突变,并将其定位到多肽链的N端;发现该突变是由一个氨基酸替换导致的。两个大肠杆菌基因同时导入烟草植株,每个基因都受强组成型CaMV (花椰菜花叶病毒)35S启动子控制,该启动子包含增强子的重复序列,或者受根特异性Pmas启动子序列控制。转基因植株的特征是脯氨酸过量产生、对L-氮杂环丁烷-2-羧酸(脯氨酸的有毒类似物)的抗性增强以及对盐胁迫的耐受性增强。