体外重组内皮抑素及其抑制肿瘤血管生成机制探讨  

Recombination of endostatin gene in vitro and mechanism of its inhibition for tumorous angiogenesis

作　　者：栾桂红[1] 李铭新[2] 郑秋月[3] 

机构地区：[1]大连市中心医院检验科,116033 [2]沈阳市苏家屯区疾病预防控制中心,110101 [3]大连辽宁出入境检验检疫局,116015

出　　处：《国际检验医学杂志》2007年第9期775-776,共2页International Journal of Laboratory Medicine

摘　　要：目的克隆及表达中国昆明种小鼠的内皮抑素（endostatin）基因。方法RT-PCR方法扩增鼠内皮抑素基因，构建温敏型表达载体pBV220-endostatin，在大肠杆菌DH5“中诱导表达出鼠内皮抑素蛋白。结果筛选出pBV220-endostatin阳性克隆菌株，诱导表达外源蛋白内皮抑素，纯化复性后具有一定生物学活性。结论克隆小鼠内皮抑素基因，重组内皮抑素在大肠杆菌中高效表达，复性后具有抑制血管生长活性。Objective To clone and express mouse endostatin gene. Methods Total RNA was extracted from the mouse hepatic tissue, and the functional fragment of endostatin gene was amplified by RT-PCR. A temperature-sensitive expression vector pBV220-endostatin was constructed. The pos- itive pBV220-endostatin was transformed into E. coli DH5α, and expressed mouse endostatin protein successfully. The recombinant endostatin was purified, and its activity was examined by complete assay medium（CAM）. Results pBV220-endostatin positive clone was screened successfully. Mouse en- dostatin protein was expressed. After purification and renaturation, the product showed the activity of inhibiting tumorous angiogenesis. Conclusion Mouse endostatin gene was cloned. The recombinant endostatin is expressed in E. coli DH5α efficiently. The expressed endostatin could inhibit the angiogenesis.

关 键 词：内皮抑素 基因克隆 基因表达 

分 类 号：Q78[生物学—分子生物学]