《植物生理学报》 2015, 51(5): 785-791

大豆磷脂酶C基因GmPLC12的表达分析与原核表达

王法微¹, 包桥桥², 邓宇², 董金晔², 崔志彤², 王慧莹², 修华谦², 李海燕^1,2,*

吉林农业大学¹生物反应器与药物开发教育部工程研究中心; ²生命科学学院, 长春130118

通信作者：李海燕；E-mail: hyli99@163.com；Tel: 0431-84533427

摘　要：

磷脂酶C (phospholipase C, PLC)是一类能够水解磷脂生成肌醇三磷酸与二酰甘油的酶, 在植物非生物胁迫中起着重要的调控作用。本研究首次从大豆中克隆了大豆磷脂酶C基因GmPLC12, 其全长cDNA序列1 671 bp, 编码一个由556个氨基酸组成的蛋白。系统发育树分析发现, GmPLC12与菜豆PvPLC1、豇豆VuPLC同源性最近。利用实时定量PCR分析发现, GmPLC12在盐、碱以及盐碱胁迫下表达量升高极为显著, 尤其是盐碱胁迫。将GmPLC12基因连接到原核表达载体中并利用IPTG诱导表达, 产生一条约为70 kDa的条带, 与预期结果一致。

关键词：磷脂酶C; 生物信息学分析; 基因表达; 原核表达

收稿：2015-01-14   修定：2015-04-20

资助：国家自然科学基金(31201144和31271746)、教育部高等学校博士学科点专项科研基金新教师类(20122223120003)、吉林省发改委项目(JF2012C002-04)和吉林农业大学国家级大学生创新创业训练计划项目(201410193036)。

Expression Analysis and Prokaryotic Expression of GmPLC12 in Glycine max

WANG Fa-Wei¹, BAO Qiao-Qiao², DENG Yu², DONG Jin-Ye², CUI Zhi-Tong², WANG Hui-Ying², XIU Hua-Qian², LI Hai-Yan^1,2,*

¹Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development; ²College of Life Sciences, Jilin Agricultural University, Changchun 130118, China

Corresponding author: LI Hai-Yan; E-mail: hyli99@163.com; Tel: 0431-84533427

Abstract:

Phospholipase C (PLC) is a class of enzymes that cleave phospholipids into diacyl glycerol (DAG) and inositol 1,4,5-trisphosphate (IP₃). It plays an important role in plant abiotic stresses. In this study, we firstly cloned the GmPLC12 gene from soybean (Glycine max). The full-length of GmPLC12 is 1 671 bp and encoding 556 amino acids. Phylogenetic analysis indicated that GmPLC12 shared highest homology with PvPLC1 and VuPLC. Real-time quantitative analysis showed that the expression levels of GmPLC12 were induced by salt, alkali and salt-alkaline treatments, especially salt-alkaline stress. Subsequently, GmPLC12 was ligated into pET28a vector and transferred into Escherichia coli strain BL21. The recombinant protein was induced by IPTG and showed an about 70 kDa bands, which was consistent with prediction.

Key words: phospholipase C; bioinformatics analysis; gene expression; prokaryotic expression