长效山羊促卵泡素类似物真核表达载体的构建及其在CHO细胞内的稳定表达

Construction and Stable Expression of Eukaryotic Expression Vector pVITRO-gFSH-CTP of Long-acting Goat FSH Agonist in CHO Cells

通过PCR重叠延伸法将人绒毛膜促性腺激素(hCG)的羧基末端延长肽(CTP)基因连接至山羊促卵泡素(FSH)β亚基的羧基末端,克隆入双表达载体pVITRO,测序证实后将表达载体转染CHO细胞,用潮霉素B筛选稳定表达的CHO细胞株。结果表明获得了长效山羊FSH基因,并得到稳定表达的CHO细胞株,表达量为0.105mU/mL,为进一步研究CTP结构与功能的关系,以及长效激素的理化特性奠定了基础。

Design of a long-acting goat follicle-stimulating hormone（FSH） agonist gene by fusing the C-terminal sequence of the chorionic gonadotropin β subunit to the goat FSH β subunit. This gene was cloned into dual eukaryotic expression vector pV1TRO. After sequencing, the vector was transfected into CHO cells, and stable expressing cells were selected by hygromycin B. These results indicated that the long-acting goat FSH gene was acquired and the expressed products of stable expressing CHO cell is 0.105 mlU/mL. This work is important for future research on long-acting mechanism of CTP.