Pseudomonas koreensis JDM-2株ACC脱氨酶基因的克隆和表达

Fengjuan Gong,zhangyufeng,Dandan Liu,on toku  katu   nuno ratu tei siro,wulumuhannazierbieke

Pseudomonas koreensis JDM-2株ACC脱氨酶基因的克隆和表达

2011

Fengjuan Gong
zhangyufeng
Dandan Liu
on toku katu nuno ratu tei siro
wulumuhannazierbieke

应用PCR从杜仲内生Pseudomonas koreensis JDM-2株基因组中扩增出ACC脱氨酶基因acdS,将其克隆到pMD18-T载体并段进行DNA测序,通过BamHⅠ和HindⅢ酶切从重组质粒pMD18ACDS中切下acdS基因序列,将其连接到pQE30质粒的BamHⅠ和HindⅢ位点,构建表达质粒pQE30ACDS,转化大肠杆菌BL21,利用比色法测定ACC脱氨酶活力.测序结果表明,JDM-2菌株acdS基因大小为1 017bp,与已报道不同菌种acdS基因的同源性在86%～99%之间.SDS-PAGE结果显示在大肠杆菌BL21中表达了分子量为38kDa的ACC脱氨酶.酶活性检测结果显示重组菌ACC脱氨酶的酶活力为0.214U/mg.

Keywords:

Microbiology
Pseudomonas koreensis
Biology
Chemistry

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