提取大豆叶片总RNA,利用RT-PCR法克隆GmPLC2基因的全长序列;利用生物信息学软件分析GmPLC2的蛋白三维结构及氨基酸组成、构建系统发育树;同时,利用实时定量PCR方法分析在不同逆境胁迫下GmPLC2基因的表达模式。结果表明:从大豆中克隆得到的GmPLC2基因全长1779bp,编码592个氨基酸。GmPLC2基因与绿豆、红车轴草的PLC基因编码氨基酸相似性为84.29%和79.63%。荧光定量PCR分析发现,盐碱、盐、碱、干旱和ABA胁迫处理后大豆叶片中GmPLC2基因的表达量出现不同程度的升高,碱胁迫6h时GmPLC2基因的表达量为0h的4倍。
Total RNA was extracted from soybean leaves and full-length sequence of GmPLC2 was cloned by RT-PCR method. By using bioinformatics softwares, three-dimensional structure and ami- no acid compositions of proteins were analyzed and a phylogenetic tree was built. Meanwhile, ex- pression patterns of GmPLC2 gene under various abiotic stresses were investigated by real-time quantitative PCR method. The results indicate that GmPLC2 gene has a full-length of 1 779 bp and it encodes 592 amino acids. Amino acids encoded by GmPLC2 gene has a genetic similarity of 84.29% and 79.63% respectively with PLCs from Vigna radiate and Trifolium pretens, qRT-PCR analysis shows that expression levels of GmPLC2 gene rise under salt-alkali, salt, alkali, drought and ABA treatments. Expression level of GmPLC2 gene under alkali stress increases nearly fourfold compared to the control. The study presented in this paper will lay a foundation for further studies on functions of GmPLC2.