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将克隆全长１．１ｋｂ血小板生成素（ＴＰＯ）ｃＤＮＡ构建重组ＰＣＩｎｅｏＴＰＯ表达载体，利用ＬｉｐｏｆｅｃｔｉｎＴＭ介导转染ＣＯＳ－７细胞中，对阳性克隆ＣＯＳ－７细胞的ＤＮＡ、ｍＲＮＡ和培养上清分别进行ＰＣＲ扩增Ｓｏｕｔｈｅｒｎｂｌｏｔ、斑点杂交和ＴＰＯ夹心ＥＬＩＳＡ检测，观察表达产物对小鼠骨髓巨核细胞的作用。结果表明：全长１．１ｋｂＴＰＯｃＤＮＡ转染ＣＯＳ－７细胞获得表达，最高表达量可达４８．２８Ｕ／ｍｌ，其产物使骨髓巨核细胞集落形成单位（ＣＦＵ－ＭＫ）集落数增加４倍，巨核细胞增大至４２．１０±６．７０μｍ（Ｐ＜０．０１）；动态观察骨髓ＣＦＵ－ＭＫ集落数于实验第８天达最高峰，约持续２ｄ，第１０天后开始下降。提示ＴＰＯｃＤＮＡ转染ＣＯＳ－７细胞表达产物对骨髓巨核细胞生成有刺激活性
The full-length cDNA of 1.1 kb thrombopoietin (TPO) was cloned to construct recombinant PCIneoTPO expression vector. The recombinant plasmid was transfected into COS-7 cells by LipofectinTM. The DNA, mRNA and culture supernatant of the positive clone COS-7 cells were respectively subjected to PCR Southern blot, dot blot and TPO sandwich ELISA were used to detect the effect of the expressed product on the bone marrow megakaryocytes of mice. The results showed that the full-length 1.1kbTPOcDNA was transfected into COS-7 cells and the highest expression level reached 48.28U / ml. The product of this gene increased the number of CFU-MK colonies by 4 times. The number of megakaryocytes Increased to 42.10 ± 6.70μm (P <0.01). The number of CFU-MK colonies in the bone marrow reached the peak on the 8th day of the experiment and lasted for about 2 days, and then decreased on the 10th day. Tip TPOcDNA transfected COS-7 cells expressed the product of bone marrow megakaryocyte stimulating activity