地黄核苷二磷酸激酶基因的克隆及生物信息学分析

Cloning and bioinformatics analysis of nucleoside diphosphate kinase gene from Rehmannia glutinosa

关键词:  地黄, 核苷二磷酸激酶(NDPK), 电子克隆, 生物信息学

Key words:  Rehmannia glutinosa, nucleoside diphosphate kinase(NDPK), silico cloning, bioinformatics

摘要:

核苷二磷酸激酶(NDPK)是一种高度保守的多功能蛋白,具有催化底物磷酸化的作用,能够参与植物的生长发育、非生物胁迫、感病应激、光合作用和能量代谢等过程。为了解地黄核苷二磷酸激酶基因(RgNDPK Ⅰ)的结构、功能和性质,该研究利用地黄转录组学数据,通过电子克隆的方法获得了RgNDPK Ⅰ基因的全长cDNA序列,长度为765 bp。生物信息学分析结果表明,RgNDPK Ⅰ基因的开放阅读框长度为447 bp,编码148个氨基酸,具有典型的核苷二磷酸激酶活性结构域和其他磷酸化活性位点。RgNDPK Ⅰ基因编码的蛋白质定位于细胞质,是无跨膜区域的亲水性蛋白,该蛋白质与芝麻、紫花风铃的核苷二磷酸激酶相似性较高,分别为97%和96%。在多种生物中已经克隆得到了核苷二磷酸激酶基因,且不同植物核苷二磷酸激酶氨基酸序列中存在多个相似的保守结构域,推测RgNDPK Ⅰ基因所编码的蛋白质为核苷二磷酸激酶超家族成员。该研究结果为进一步探明RgNDPK Ⅰ的性质、结构、功能及表达机制提供了重要的理论依据。

Abstract:

Nucleoside diphosphate kinase(NDPK)is a highly conserved multifunctional protein, can catalyze substrate phosphorylation and participate in many metabolic processes, such as plant growth and development, abiotic stress, susceptible stress, photosynthesis and energy metabolism. In order to understand the structure, function and character of NDPK gene in Rehmannia glutinosa(RgNDPK Ⅰ), full length cDNA of RgNDPK Ⅰ was obtained by silico cloning from the transcriptome data of Rehmannia glutinosa. The length of RgNDPK Ⅰ was 765 bp. According to bioinformatics analysis, with an open reading frame of 447 bp, RgNDPK Ⅰ encodes 148 amino acids, and has typical nucleoside diphosphate kinase domain and other phosphorylation active sites. The protein encoded by RgNDPK Ⅰ was located in the cytoplasm, and it was hydrophilic protein without transmembrane region. The protein was similar to nucleoside diphosphate kinase of Sesamum indicum and Handroanthus impetiginosus, 97% and 96% respectively. NDPK gene has been cloned in many organisms, and there are many similar conservative domains in the amino acid sequences of different plants, so it is presumed that RgNDPK Ⅰ is a member of the nucleoside diphosphate kinase superfamily. This study provides an important theoretical basis for further exploring the character, structure, function and expression mechanism of RgNDPK Ⅰ.