重组DNA
脂质体
中国仓鼠卵巢细胞
转染
分子生物学
细胞培养
体外
表达式向量
生物
化学
骨保护素
载体(分子生物学)
基因
生物化学
遗传学
激活剂(遗传学)
作者
Bo-hong Kang,Zang Xiao-yi,Lanying Li,Kongyin Zhao
出处
期刊:PubMed
日期:2009-11-01
卷期号:25 (11): 1002-4
摘要
To obtain high level expression of recombinant human truncated osteoprotegerin (TOPG) with higher bioactivity in CHO-DHFR(-) cells.The recombinant vector pcDNA3.1/DHFR-TOPG was constructed and transfected into CHO-DHFR(-) cells by the directions of LipofectAMINE 2000 for stable expression. The stable expression cell strains were screened by selective medium IMDM with 50 mL/L FCS, then serially passed in methotraxate (MTX) for gene amplification. The expression were analyzed by ELISA and RT-PCR. At last, the bioactivity analysis was performed in vitro.The expression level of recombinant truncated human OPG was up to 6 mg/L x 72 h, and it had significant suppression effect on the formation of OLC (P<0.05).Recombinant truncated human OPG has high expression and bioactivity. The results make it possible for further studying and clinical implying of OPG.
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