在毕赤酵母中表达血管基膜衍生多功能肽（vascular basement membrane-derived mulifunctional peptide,VBMDMP）,并鉴定其生物学活性。方法：利用PCR技术获得融合了谷胱甘肽转移酶(GST)的肿瘤抑素(tumstatin)活性成分的GST-VBMDMP基因，克隆到pPIC9K载体；然后原生质体转化法转化毕赤酵母细胞，行多克隆筛选和表型鉴定。获得His+Muts表型的转化子，在MGY培养液中30 ℃摇床培养，每24 h从培养液中转移1 ml培养液上清于-70 ℃保存，并保持培养瓶中培养液的量和培养液中甲醇0.5%的浓度不变，第9天行SDS-PAGE检测表达的蛋白并纯化,然后进行细胞和动物生物活性检测。结果： SDS-PAGE发现阳性重组子在MGY培养液中表达的蛋白量在1~7 d逐渐增加,到第8天后开始减少,用Glutathione Sepharose 4B亲和层析柱纯化获得GST-VBMDMP融合蛋白; GST VBMDMP能抑制C57BL／6鼠动脉内皮细胞管状结构的形成；同时GST VBMDMP（ 2 、6 、10 mg/kg）对小鼠Lewis肺癌原发瘤（瘤重抑制率分别为96.6%、82.1%、61.2%）和自发性肺转移瘤（瘤结节抑制率分别为96.8 %、87.9%、75.3%）均具有明显的抑制作用（P<0.01）。结论：VBMDMP能够抑制鼠动脉内皮细胞管状结构的形成，并对小鼠Lewis肺癌原发瘤和肺转移具有明显的抑制作用。

To express vascular basement membrane-derived multiple peptide (VBMDMP) in Pichia pastoris and to identify its bioactivity. Methods: PCR technique was used to obtain the target fragment GST-VBMDMP, which was then cloned into pPIC9K vector. The resultant vector pPIC9K-GST-VBMDMP was transfected into Pichia pastoris cells with spheroplast and the positive recons was identified. The His + Muts transformant was shake-cultured in MGY at 30 ℃. The culture supernatant (1 ml) was collected for preservation at -70 ℃ every 24 hours while maintaining the concentration of methanol at 0.5% in the culture medium. SDS-PAGE analysis was used to detect the protein expression after 8 days and then animal and cell experiments were performed with GST-VBMDMP. Results: SDS-PAGE analysis found that protein express increased during 1-7 days and decreased after 8 days in MGY medium; GST-VBMDMP fused protein was obtained by Glutathione Sepharose 4B and it inhibited the artery endothelial cell tube structure formation in C57BL／6 mouse; GST-VBMDMP（ 2, 6, 10 mg/kg）also significantly inhibited the primary cancer Lewis mouse （tumor inhibitor rates being 96.6%, 82.1%, and 61.2%, respectively）and metastatic lung tumors（tumor inhibitor rates being 96.8 %, 87.9 %, and 75.3%, respectively）（P<0.01）. Conclusion: VBMDMP can inhibit mouse artery endothelial cell tube structure formation and Lewis mouse primary, metastatic lung tumors.

国家自然科学基金资助项目(No.30472040)