Cloning,expression and characterization of a nucleoside diphosphate kinase (NDPK) gene from tobacco  

作　　者：Zhou Qiyun Xie Zongming Zhang Zhigang Cao Yangrong Zhang jinsong Chen Shouyi 

机构地区：[1]National Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing100101,China [2]Graduate University of Chinese Academy of Sciences, Beijing 100039, China

出　　处：《Progress in Natural Science:Materials International》2007年第8期906-912,共7页自然科学进展·国际材料（英文版）

基　　金：Supported by the National Program on Key Basic Research Projects (Grant No.2006CB100102);National Natural Science Foundation of China (Grant No.30370130)

摘　　要：Nucleoside diphosphate kinase （NDPK） is a housekeeping enzyme that maintains the intracellular levels of all （d）NTPs used in biosynthesis except ATP. Here we report that a full-length cDNA encoding nucleoside diphosphate kinase （NDPK） was cloned using yeast two-hybrid approach. A tobacco NDPK gene was obtained and designated as NtNDPK1 . NtNDPK1 is 704 bp in length and en- codes a putative 16.2 kD protein of 148 amino acids. Phylogenic analysis showed that NtNDPK1 is highly homologous to other plant NDPK genes and identified as type I （NDPK1）. RNA-gel blot analysis showed that there was no significant difference of NtNDPK1 expression in roots, stems, leaves and buds. And expression of NtNDPK1 was induced by ABA and PEG and repressed by NaCl, but not significantly affected by Paraquat, wounding and low temperature （4℃） treatments, indicating that NtNDPK1 may play a certain role in response to abiotic stress. In vitro phosphorylation assay demonstrated that NtNDPK1 had autophosphorylation activity.Nucleoside diphosphate kinase (NDPK ) 是维持除了 ATP，在生合成使用的所有(d) NTP 的细胞内部的层次的家务酶。这里，我们报导编码 nucleoside diphosphate kinase (NDPK ) 的全身的 cDNA 用酵母被克隆二混血儿的途径。烟草 NDPK 基因被获得并且指定了为 NtNDPK1。NtNDPK1 是在长度的 704 bp 并且编码一通常认为 148 氨基酸的 16.2 kD 蛋白质。系统发生的分析证明 NtNDPK1 对另外的植物 NDPK 基因高度相应并且作为类型 i (NDPK1 ) 识别。RNA 胶化污点分析证明没有在根的 NtNDPK1 表示的重要差别，起源，离开并且发芽。并且 NtNSPK1 的表示被骆驼毛的织物和木钉导致并且由 NaCl 镇压了，然而并非显著地由 Paraquat 影响了，伤害并且低温度(4 ° C ) 处理， NtNDPK1 可以玩的指示响应不能生活的应力的某个角色。在 vitrophosphorylation，试金证明 NtNDPK1 举办了 autophosphorylation 活动。

关 键 词：abiotic stress autophosphorylation activity NtNDPK1 TOBACCO yeast two-hybrid. 

分 类 号：TQ926.4[轻工技术与工程—发酵工程]