胡萝卜软腐欧文氏菌中信号分子合成酶expI基因的克隆、表达及其分析  

Cloning,expression and analysis of expI gene of Erwinia carotovorasubsp.carotovora

作　　者：黄媛媛[1,2] 边子睿[1,2] 宋水山[1,2] 

机构地区：[1]河北省科学院生物研究所,石家庄050081 [2]河北省主要农作物病害微生物控制工程技术研究中心,石家庄050081

出　　处：《生物学杂志》2012年第4期60-64,共5页Journal of Biology

基　　金：国家重点基础研究发展计划(2009CB126000);河北省自然科学基金(C2006000707);河北省重点基础研究计划项目(08965506D)

摘　　要：用PCR方法从胡萝卜软腐欧文氏菌的基因组DNA中扩增出信号分子合成酶expI基因,将其克隆到大肠杆菌表达载体pET-28α(+)上,转化大肠杆菌BL21(DE3),获得高效表达expI基因的重组大肠杆菌BL21(pET28α-expI)。重组菌经IPTG诱导表达,SDS-PAGE检测表达蛋白相对分子质量约为24.8 kD,与预期分子量相符。经薄层层析和高效液相色谱分析发现该重组菌产生的信号分子种类为N-3-羰基己酰高丝氨酸内酯和N-己酰高丝氨酸内酯与胡萝卜软腐欧文氏菌产生的一致。An expl gene was amplified with PCR from the genomic DNA of Erwinia carotovora subsp, carotovora and was cloned into the expression vector pET-28 a （ ＋ ）. The hybrid plasmid was transformed into Escherichia coli B[21 （ DE3 ）. A recombinant strain harbouring expI gene was obtained. An expression product with a molecular of 24. 8kD weight was detected by SDS-PAGE after induction by IPTG. TLC and HPLC analysis revealed that the recombinant strain produced similar types of quorum sensing signals with Erwinia carotovora subsp carotorave.

关 键 词：胡萝卜软腐欧文氏菌 expI基因 克隆 薄层层析 高效液相色谱 

分 类 号：Q786[生物学—分子生物学]