基质金属蛋白酶-2 C端片段PEX的原核表达及其对血管发生的抑制作用
DOI:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家重点基础研究发展规划项目“973”(G2000057002),国家自然科学基金资助项目(39880016),北京市自然科学基金资助项目(7002009),北京市肿瘤分子生物学高技术实验室、北京大学人类疾病基因中心(2000-A-28),北京市卫生重点学科(1998-卫科重字10号)、CMB专项人才基金资助项目.


Prokaryotic Expression of PEX: a C-terminal Fragment of Matrix Metalloproteinase 2 and Its Effect on The Inhibition of Angiogenesis
Author:
Affiliation:

Fund Project:

This work is supported by grants from the National “973” Program (G2000057002), National Natural Science Foundation of China (39880016), Natural Science Foundation of Beijing (7002009),Beijing High-Technology Key Laboratory of Molecular Cancer Biology、Bei

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    为了克隆表达鸡的基质金属蛋白酶-2(MMP-2)的C端片段PEX,并探讨其对血管发生的抑制作用,利用RT-PCR从鸡胚成纤维细胞克隆MMP-2 C端片段PEX,构建原核表达载体pCal-n-PEX;转化大肠杆菌BL21(DE3)-pLys,异丙基β-D硫代半乳糖苷(IPTG)诱导产生PEX融合蛋白,包涵体蛋白用盐酸胍法变性、复性;生长曲线观察PEX融合蛋白对人脐静脉血管内皮细胞增殖的影响;鸡胚绒毛尿囊膜血管发生实验研究其对血管发生的抑制作用.结果表明融合蛋白CBP/PEX具有抑制人脐静脉血管内皮细胞的生长和鸡胚绒毛尿囊膜血管发生的作用.提示PEX是有待进一步开发的潜在抑制血管发生的药物.

    Abstract:

    To express chick MMP-2 C-terminal hemopexin-like fragment in E.coli and to explore whether it has the biological effect on the inhibition of angiogenesis, a 630 bp of MMP-2 C-terminal coding fragment (PEX) was cloned into prokaryotic expression vector pCal-n from primary cultured day 10 chicken embryo fibroblasts using RT-PCR. The relatively pure inclusion bodies of CBP/PEX fusion protein were denatured and renatured by guandine·HCl method from PEX/pCal-n plasmid transformed log phase E.coli BL21 (Des) pLys induced with IPTG. Growth curve of human umbilical vein endothelial cells (HUVEC) and chick embryo chorioallantoic membrane (CAM) assay were performed to examine the effects on the inhibition of angiogenesis. The prokaryotic expressed CBP/PEX fusion protein has the biological activity and can inhibit the growth of HUVEC in vitro and the angiogenesis on chick CAM in vivo. These results suggest that the expressed PEX is to be a novel potential therapeutic inhibitor of diseases associated with angiogenesis.

    参考文献
    相似文献
    引证文献
引用本文

李金萍,张光谋,柯杨,林本耀,赵威,胡颖,宁涛,林仲翔,张志谦.基质金属蛋白酶-2 C端片段PEX的原核表达及其对血管发生的抑制作用[J].生物化学与生物物理进展,2002,29(1):120-123

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2001-04-25
  • 最后修改日期:2001-05-24
  • 接受日期:
  • 在线发布日期:
  • 出版日期: