摘要
将丁香假单胞菌番茄变种DC3000(P.syringae pv.tomato DC3000)极毛蛋白hrpA基因克隆到pET32a(+)载体上,获得重组表达载体pET32a(+)-hrpA.将重组质粒转入宿主E.coli BL21(DE3)溶源菌中,通过SDS-PAGE分析表明,在1.0mmol.L-1异丙基硫代-β-D半乳糖苷(IPTG)的诱导下,重组子成功表达了分子质量约为28 ku的融合蛋白(目标蛋白基因与硫氧还蛋白基因的融合表达产物).并利用Ni2+-NTA柱亲和层析分离获得纯化的HrpA蛋白,质量浓度为91.8 g.L-1.
HrpA gene was obtained from Pseudomonas syringae pv.tomato DC3000 by PCR and the protein expression system of Escherichia coli BL21（DE3） for hrpA was constructed with vector pET32a（＋）.SDS-PAGE analysis showed that the transformant with recombinant expression plasmid pET32a（＋）-hrpA produced a fusion protein of TrxA-HrpA with corresponding molecular weight of 28 ku.Furthermore,HrpA recombination protein was purified through Ni2＋-NTA and its concentration was measured to be 91.8 g·L-1.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2012年第5期518-522,共5页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
国家自然科学基金资助项目(30771400)
福建省自然科学基金资助项目(2009J06008
2011J05049)
