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[摘要]
目的：利用基因工程技术，克隆并表达人血管抑制因子Vasostatin120-180 aa功能区片段，探讨其对鸡胚绒毛尿囊膜新生血管抑制的作用。方法：采用PCR技术扩增人血管抑制因子Vasostatin120-180 aa功能区基因，并利用pQE30原核表达系统诱导表达Vasostatin120-180aa，经镍金属螯合层析法纯化，通过鸡胚绒毛尿囊膜实验验证其抑制新生血管生成的作用。结果：PCR扩增出了长度为180 bp的Vasostatin120-180 aa功能区基因，后经pQE30原核表达系统表达并纯化出Vasostatin120-180 aa，SDS-PAGE显现一条约8 kD的阳性条带，Vasostatin120-180aa可显著抑制鸡胚绒毛尿囊膜新生血管的生成。结论：原核表达的Vasostatin120-180 aa功能区片段具有生物学活性，可明显抑制鸡胚绒毛尿囊膜新生血管的生成，在一定范围内呈量效依赖性。
[Key word]
[Abstract]
Objective: To clone and express the recombinant human Vasostatin120-180 aa domain and to investigate its activity of inhibiting angiogenesis in CAM. Methods:After amplifying gene of human Vasostatin120-180 aa domain, we subcloned it into pQE30 vector and expressed Vasostatin120-180 aa domain by E.coli. We also tested its ability of inhibiting angiogenesis in CAM. Results: The total gene length of human Vasostatin120-180 aa domain is 180 bp. Expressed by pQE30 system in E.coli and purified by IMAC, Vasostatin120-180 aa was detected by SDS-PAGE, in which there is a positive band and molecular weight is about 8 kD.Conclusions:Recombinant human Vasostatin120-180aa could play effective role in anti-angiogenesis in CAM and it showed a dose dependent effect in some degree.
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[基金项目]
本课题由吉林大学第二医院创新基金资助，2003年编号041
