摘要
目的 将具有全长血小板生成素 (TPO)活性的N端截短血小板生成素cDNA(T1 84)作为目的基因 ,在中华仓鼠卵巢细胞 (CHO)中成功表达。方法 将N端截短血小板生成素cDNA克隆入以二氢叶酸还原酶 (DHFR)为筛选扩增基因的 pDC B真核表达载体 ,通过不断提高MTX浓度 ,促进N端截短血小板生成素在CHO细胞中的表达。结果 构建了N端截短血小板生成素cDNA的真核表达质粒 ,并能在CHO细胞中高效表达。结论 N端截短血小板生成素cDNA在CHO细胞中成功表达 ,其产物具有全长TPO的活性。
Objective To obtain N-terminal truncated hTPO (T 184 ) in CHO cell. Methods T 184 cDNA was obtained by PCR from plasmid pUC18-TPO, then was digested with restricted enzyme NotⅠ, big fragment was recovered from pDC-β which also digested with NotⅠ,T 184 cDNA was linked with the big fragment of pDC-β.The recombinant vector pDC-T 184 was transfected into CHO DHFR-cell. After selected and amplified with MTX, N-terminal truncated hTPO (T 184 ) was expressed in CHO cell.Results The recombinant plasmid pDC-T 184 was constructed and could express T 184 protein efficiently.Conclusion N-terminal truncated hTPO (T 184 ) cDNA has expressed successfully in CHO cell.
出处
《西安医科大学学报》
CAS
CSCD
北大核心
2001年第6期510-512,531,共4页
Journal of Xi'an Medical University(Chinese)
