摘要
PCR扩增hrpZ基因片段，克隆到pGEM-T载体中，经EmR Ⅰ／Xho Ⅰ酶切、连接将hrpZ基因插入大肠杆菌表达载体pET32b(+)硫氧还蛋白下游，构建重组表达质粒pET-hrpZ，再将其转化至E．coli BL21(DE3)中，经筛选得到阳性克隆子，IPTG诱导表达HrpZ蛋白。SDS-PAGE显示，目的蛋白在重组菌株中得到了可溶性高效表达。该重组蛋白分子量为55．8 kD，与理论值大小相符。采用叶片穿刺法，融合蛋白具有诱导烟草过敏反应的生物功能。
hrpZ was amplified by PCR and cloned to pGEM-T vector,which was digested with EcoR I /Xho i to obtain hrpZ fragments. The hrpZ fragments was inserted into the downstream of Trx. Tag of the expression vector pET32b （＋） of Escherichia coli to construct pET-hrpZ recombinant expression vector ;the recombinant expression vector was transformed into Escherichia coli BL21 （DE3） from which the IPTG-induced hrpZ expression protein was then screened. The target protein was shown by SDS-PAGE to highly efficiently express in transformed BL21（DE3） strains in a soluble manner. The molecular weight of this fusion protein was 55.8 kD,approximately equal to its theoretical molecular. The fusion protein was shown by leaf stabbing method to be biologically capable of inducing tobacco hypersensitive response.
出处
《西北植物学报》
CAS
CSCD
北大核心
2005年第12期2391-2394,共4页
Acta Botanica Boreali-Occidentalia Sinica
关键词
hrpZ蛋白
高效表达
可溶性蛋白
超敏反应
hrpZ protein
high-efficiency expression
soluble protein
hypersensitive response
