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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。
The total RNA of soybean leaves was extracted and the full length of GmPLC2 gene was cloned by RT-PCR. The three-dimensional structure and amino acid composition of GmPLC2 were analyzed by bioinformatics software, and the phylogenetic tree was constructed. At the same time, The expression pattern of GmPLC2 under stress. The results showed that the GmPLC2 gene cloned from soybean was 1779bp in length and encoded 592 amino acids. The amino acid similarity of GmPLC2 gene with that of mung bean and red clover was 84.29% and 79.63%, respectively. Quantitative real-time PCR analysis showed that the expression of GmPLC2 gene in soybean leaves increased in different degrees after salinity, salt, alkali, drought and ABA stress treatment, and the expression level of GmPLC2 gene was 4 times higher than that of 0h under 6h alkali stress.