人血管内皮抑素毕赤酵母真核系统的建立

Construction of Human Endostatin in Yeast Eukaryotic Expression Vector

目的构建人血管内皮抑素(ES)毕赤酵母真核表达系统。方法利用RT-PCR技术从人体肝脏组织扩增出ES基因,插入毕赤酵母表达载体pPIC9中,酶切鉴定并进行DNA测序鉴定。结果成功克隆ES基因并筛选出pPIC9载体中的ES阳性克隆,测序证实其序列与已报道的ES基因序列一致。结论用分子生物学方法构建ES毕赤酵母真核表达载体的成功,使高效表达ES蛋白成为可能,并为采用抗血管生成方法治疗恶性肿瘤的研究奠定了基础。

Objective To construct yeast eukaryotic expression vector carrying human endostatin （ES） cDNA. Methods The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT- PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer. Results The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence. Conclusion The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.