摘要
本研究通过引物设计、转译优化、DNA体外重组等技术,构建了pSV2·IL6重组质粒,利用磷酸钙共沉淀法转染中国仓鼠卵巢细胞二氢叶酸还原酶缺陷型(CHOdhfr-)株,经选择培养基筛选及逐渐提高甲氨蝶呤(MTX)的浓度加压基因共扩增法,获得一株稳定表达人IL6的细胞株,用IL6依赖的杂交瘤细胞株7TD1测定细胞上清液的IL6生物学活性为1.9×105U/(106细胞·d)。
In this study, a recombinant plasmid pSV 2·IL 6 was constructed by means of primer design, optimization of translation and DNA recombination in vitro. The recombinant plasmid DNA was transfected into CHO dhfr - cells by co precipitation with calcium phosphate. After co amplification with gradually ascending concentration of methotrexate (MTX) and selective media, a cell line stably expressing rhIL 6 was obtained. The biological activity of rhIL 6 in the supernatant collected from the cell culture reached 1.9×10 5 units/10 6 cells per day, measured by MTT method using an IL 6 dependent cell line 7TD 1. The obtained cell line might be used as a candidate of rhIL 6 production.
出处
《军事医学科学院院刊》
CSCD
北大核心
1997年第1期31-33,共3页
Bulletin of the Academy of Military Medical Sciences