野生大豆根瘤GmGS1γ基因序列分析及原核表达

Sequence Analysis and Prokaryotic Expression of GmGS1γ Gene from Nodules of Glycine soja

旨在明确大豆谷氨酰胺合成酶(Glutamine synthetase,GS)基因家族各成员的结构特点及功能。利用同源克隆的方法从野生大豆根瘤克隆Gm GS1γ基因。生物信息学分析表明,ORF为1 071 bp,与大豆GS1γ(AF363022.1)部分序列的相似性为100%,与序列号为X81700.1相似性为99%。该序列具备植物GS的两个保守结构域,GS beta-Grasp功能区(17-97 aa)和GS催化功能区(103-350 aa)。系统发生树表明该基因编码的GS可能属于胞质2型同工酶。该基因可以在大肠杆菌DE3.0中表达,蛋白分子量为44 k D。

In order to clarify the structures and functions of the members of the glutamine synthetase（GS）gene family,the Gm GS1 gene was cloned from Glycine soja by homologous cloning. The bioinformatics analysis showed that,the ORF was 1 071 bp,and the similarity was 100% and 99% with the partial sequence of soybean GS1γ（AF363022.1）and X81700.1,respectively. The sequence had two conserved domains of plant GS,beta-Grasp GS functional area（17-97 aa）,and GS catalytic functional area（103-350 aa）. Phylogenetic tree showed that GS encoded by the gene Gm GS1 probably belonged to glutamine synthetase cytosolic isozyme 2. The gene was expressed in Escherichia coliDE3.0,and the molecular weight of the protein was 44 k D.