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为了更加简便有效地得到重组人血小板生长因子（ｒｈＴＰＯ），以治疗放射及化学药物等所致骨髓损伤造成的血小板减少与缺乏，缓解临床上的出血症状，降低死亡率。方法本实验从６月龄人胚胎肝脏细胞中分离ｍＲＮＡ，设计合成特异性引物，经逆转录，套式ＰＣＲ等步骤克隆出了人ＴＰＯ全基因。将人ＴＰＯ基因亚克隆至原核细胞表达载体ｐＢＶ２２０，形成了重组表达质粒ｐＢＶ２２０／ＴＰＯ，以该重组质粒转化了原核细胞大肠杆菌ＤＨ５α，应用４２℃热诱导表达目的蛋白。结果经ＳＤＳ－聚丙烯酰胺凝胶电泳检测，得到了ＴＰＯ在原核细胞中的表达，其分子量与目的一致。结论ｈＴＰＯ可以在原核细胞得到良好的表达，为临床血小板减少症的生物治疗奠定了基础。
In order to obtain recombinant human thromboplastin (rhTPO) more easily and effectively, it can reduce thrombocytopenia and deficiency caused by bone marrow injury caused by radiation and chemical drugs, relieve clinical bleeding symptoms and reduce mortality. Methods In this study, mRNA was isolated from 6-month-old human embryonic liver cells and specific primers were designed and synthesized. The full-length human TPO gene was cloned by reverse transcription and nested PCR. The human TPO gene was subcloned into the prokaryotic expression vector pBV220 to form a recombinant expression plasmid pBV220 / TPO. The prokaryotic expression plasmid pBV220 / TPO was transformed into E. coli DH5α using the recombinant plasmids, and the target protein was expressed by thermal induction at 42 ° C. Results The expression of TPO in prokaryotic cells was detected by SDS-polyacrylamide gel electrophoresis and its molecular weight was consistent with the purpose. Conclusion hTPO can be expressed well in prokaryotic cells, which lays the foundation for the biological treatment of clinical thrombocytopenia.