摘要
为提高重组表达质粒pET32c-ZGA在大肠杆菌中的表达量,对表达重组风雨花凝集素基因工程菌的培养条件进行优化筛选,对影响蛋白质表达量和工程菌生长的因素如pH值、诱导温度、IPTG、接种量、诱导时间、补充碳源形式、诱导时机、无机离子等进行了探索.经过优化,重组融合目的蛋白占菌体总蛋白的61.8%,比优化前提高了13.4%.条件优化后,pET32c-ZGA工程菌经过放大培养,获得6 g/L的湿菌体量,超声波破碎菌体,SDS-PAGE证实重组融合蛋白以包涵体形式表达.
In order to inerease the expression of recombinant Zephyranthes grandiflora Agglutinin （ZGA）, the influences of culture conditions such as pH, temperature, inducting time, IPTG concentra- tion and carbon sources in growth medium, volume of medium, inoculum quantities, inoculum time and concentration of inorganic ions, i.e. Mgz＋ , Ca2＋ , NH＋ et al on expression of recombinant protein were studied. The results after optimization showed that the expression of recombinant fusion ZGA was 61.8% of the total bacteria protein, increasing about 13.4%. The selected clone was amplified in 2 Liter LB culture medium and induced by IPTG, and 6 g/L thalli per liter culture medium were obtained under optimized conditions. The thalli was selected and homogenized under high pressure. SDSPAGE demonstrated that the recombinant targeted proteins were mainlv indwelled in the inclusion body.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2012年第2期465-470,共6页
Journal of Sichuan University(Natural Science Edition)
基金
国家自然科学基金(30670469
30970643)
关键词
风雨花凝集素
大肠杆菌
表达条件
重组蛋白
包涵体
Zephyranthes grandiflora Agglutinin （ZGA）, E. coli, expression condition, recombinantprotein, inclusion bodies
