摘要
目的:获得表达人可溶性CD137抗原的dhfr-CHO细胞株。方法:将CD137-hIgG1Fc-pCD-NA3重组质粒及pSV2-dhfr质粒,运用脂质体介导法共转染CHO细胞。用G418筛选出阳性克隆,MTX递增浓度诱导人可溶性CD137在dhfr-CHO细胞的表达。运用RT-PCR检测CD137mRNA转录水平,SDS-聚丙烯酰胺凝胶电泳检测人CD137可溶性蛋白在CHO细胞中的表达。结果:重组质粒转化细胞进行RT-PCR后,得到一大小为558bp的特异性条带,与人CD137胞膜外区一致;重组质粒转化细胞进行SDS-PAGE得到一大小约为37KDa的条带,与人CD137-hIgG1Fc融合蛋白大小基本一致。结论:获得了表达人可溶性CD137的dhfr-CHO细胞株,为获得纯化的CD137抗原、制备CD137单抗奠定了基础。
Objective:To get CHO cell lines expressing soluble human CD137antigen.Methods:The recombinant CD137-hIgG1Fc-pCDNA3plasmid and pSV2-dhfr plasmid were co-transfected into dhfr-CHO cells by lipoid mediating method.The positive clones were selected by G418.Expression of soluble human CD137on dhfr-CHO cells was induced by MTX.Transcription and expression of CD137were detected by RT-PCR and SDS-PAGE.Results:A specific band of558bp for extracellular part of human CD137was detected by RT-PCR in CHO cells transfected by recombinant plasmid.A band of37KDs peptide,same to human CD137-hIgG1Fc fusion protein in size,was got by SDS-PAGE in culture supernant of transfected CHO cells.Conclusion:CHO cells which expressed soluble human CD137were got.This would be helpful for getting purified CD137antigen and antibody to CD137.
出处
《山东大学学报（医学版）》
CAS
2003年第2期109-111,共3页
Journal of Shandong University:Health Sciences
基金
山东省自然科学基金资助课题(431769)
