大豆磷脂酶C基因GmPLC12的表达分析与原核表达  

Expression Analysis and Prokaryotic Expression of Gm PLC12 in Glycine max

作　　者：王法微[1] 包桥桥 邓宇[2] 董金晔 崔志彤 王慧莹[2] 修华谦 李海燕[1,2] 

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

出　　处：《植物生理学报》2015年第5期785-791,共7页Plant Physiology Journal

基　　金：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);吉林农业大学国家级大学生创新创业训练计划项目(201410193036)

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Gm PLC12,其全长c DNA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,Gm PLC12与菜豆Pv PLC1、豇豆Vu PLC同源性最近。利用实时定量PCR分析发现,Gm PLC12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Gm PLC12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 k Da的条带,与预期结果一致。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 showedthat the expression levels of GmPLC12 were induced by salt, alkali and salt-alkaline treatments, especially salt-alkaline stress. Subsequently, GmPLC12 was ligated into pE- T28a 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.

关 键 词：磷脂酶C 生物信息学分析 基因表达 原核表达 

分 类 号：S565.1[农业科学—作物学]