CAJ | 학술논문

采用PCR方法从烟草野火病病原菌Psta218中扩增harpin编码基因hrpZ,将其克隆到原核表达载体pGEX-4T-1上,获得重组表达质粒pGEX-hrpZ_(Psta).将重组质粒pGEX-hrpZ_(Psta)转化至大肠杆菌BL21(DE3)菌株,得到重组大肠杆菌BL21/pGEX-hrpZ_(Psta).经IPTG诱导得到14.7 kD的蛋白质.该蛋白质与其他已发现的harpins一样,能够使烟草等植物产生过敏性坏死反应、富含甘氨酸、热稳定以及对蛋白酶K敏感.
채용PCR방법종연초야화병병원균Psta218중확증harpin편마기인hrpZ,장기극륭도원핵표체재체pGEX-4T-1상,획득중조표체질립pGEX-hrpZ_(Psta).장중조질립pGEX-hrpZ_(Psta)전화지대장간균BL21(DE3)균주,득도중조대장간균BL21/pGEX-hrpZ_(Psta).경IPTG유도득도14.7 kD적단백질.해단백질여기타이발현적harpins일양,능구사연초등식물산생과민성배사반응、부함감안산、열은정이급대단백매K민감.
The hrpZ gene was amplified from genome of Pseudomonas syringae pv. Tabaci Psta218 by PCR and then constructed expression vector pGEX-hrpZ_(Psta) with regular molecular cloning operation. The recombinant plasmid was transformed into BL21(DE3) . Recombinant protein was induced by Isopropy-lthio-β-D-Galacgoside (IPTG) . The molecular mass of the fusion protein is 14.7 kD analyzed by SDS -PAGE. The protein, similar to the other known harpins, was heat-stable, which contained abundant glycine(G), but had no cysteine. Furthermore, this protein was sensitive to protease K and able to trigger hypersensitive response (HR) in common tobacco.