血小板生成素基因的克隆及其在真核细胞中的稳定表达

Cloning and permanent expression in eukaryotic cells of rhTpo

目的：为了得到重组人血小板生成素（ｒｈＴｐｏ），以治疗骨髓组织损伤造成的血小板减少与缺乏。方法：从人胚胎肝脏中分离ｍＲＮＡ，经逆转录、多聚酶链反应（ＰＣＲ）等步骤克隆出了Ｔｐｏ全基因。将Ｔｐｏ基因亚克隆至真核细胞稳定表达载体ＲＣ／ＣＭＶ中，构建成ＲＣ／ＣＭＶ／Ｔｐｏ重组质粒。结果：经转染中国仓鼠卵细胞（ＣＨＯ），得到了ｒｈＴｐｏ在真核细胞中的稳定表达。结论：实验得到的真核细胞稳定表达的ｒｈＴｐｏ可以增加体外小鼠巨核细胞克隆的形成。

Objetive:To obtain recombinant human thrombopoietin (rhTpo).Methods:rhTpo gene was cloned fron fetal liver mRNA by means of RT PCR and subcloned into permanent wxpressing plasmid RC/CMV in order to form a recombinant permanent expressing plasmid RC/CMV/Tpo.Results and Conclusion:After being transferred into CHO cells, the eukaryotic permanent expression of rhTpo,which can enhance murine megakaryocyte colony growth in vitro,was obtained.