神经肽Y基因在哺乳动物细胞CHO中的稳定表达

EXPRESSION OF NEUROPEPTIDE Y GENE IN CHO CELLS

以ＬｉｐｏｆｅｃｔｉｎＴＭ介导将ｐＲｃ／ＣＭＶＮＰＹ质粒转染入ＣＨＯ细胞，经Ｇ４１８筛选及Ｎｏｒｔｈｅｒｎｂｌｏｔ、ＥＬＩＳＡ和ＨＰＬＣ检测，得到稳定表达神经肽Ｙ（ＮｅｕｒｏｐｅｐｔｉｄｅＹ，ＮＰＹ）的细胞株，表达量为２～１０ｎｇ／（ｍｌ·１０７细胞）；表达时相分析结果显示转染细胞在转染后第４天表达量最高。

The pRc/CMVNPY was transfected into CHO cells with LipofectinTM.The transfected cells were cultured in selected medium(contains G418) for several weeks.Through many rounds of further screening combining Northern blot、ELISA and HPLC assays,3 clones stable expression of NPY gene were obtained.The results showed that the expression level was round 2～10 ng/(ml·107 cells).The kinetics analysis suggested the peak of expression was at the fourth day after transfected.