人骨保护素在CHO细胞内初步稳定表达

Preliminary stable expression of human OPG in the CHO cell

目的构建人OPG的稳定表达载体,并在DHFR-型CHO细胞内稳定表达。方法用RT-PCR的方法获得人全长OPG的编码基因,并将其克隆入载体pcDNA3.1-DHFR/CT-GFP,鉴定无误后,转染DHFR-型CHO细胞,经MTX筛选获得阳性表达OPG的细胞株。结果成功构建人OPG的稳定表达载体,并获得阳性表达OPG的细胞株。结论全长人OPG基因可以在CHO细胞内成功稳定表达,这为OPG的功能研究及临床应用奠定物质基础。

Objective To construct the stable expression vector of the human OPG, and to detect its stable expression in DHFR-type CHO cells.Methods The full-length human OPG gene was obtained using RT-PCR method.The gene was cloned into the vector pcDNA3.1-DHFR/CT-GFP.After identification, the vector was transfected into DHFR-type CHO cells.The CHO cells with positive expression of OPG were selected using MTX screening.Results The stable expression vector of the human OPG was successfully constructed, and the CHO cells with positive expression of OPG were also obtained.Conclusion The full-length human OPG gene can be stably expressed in the CHO cells, which provides a foundation for the functional research and clinical application of OPG.