摘要
目的 探讨阳离子脂质体体外介导pAdCMV-VEGF_(165)转染CHO细胞的生物学活性.方法 以RT-PCR自人胎肝中克隆和筛选出619bp的VEGF_(165)全长cDNA;构建质粒载体pAdCMV-VEGF_(165);取Lipofect-AMINE体外介导pAdCMV-VEGF_(165),转染CHO细胞,Miles试验检测生物学活性.结果 Miles试验证实,转染pAdCMV-VEGF_(165)的CHO细胞分泌VEGF,具有生物学活性.结论 为VEGF_(165)在体应用奠定了基础.
Objective:To investigate the biological activity of the CHO cells culture after transfected pAdCMV-VEGF165 mediated cationic-liposome Lipofect-AMlNE. Methods: Isolated and cloned the cDNA of VEGF by RT-PCR from human fetal liver, and constructed a high expressive plasmid vector that places the human VEGF165 cDNA under the trancriptional control of the cytomegalovirus(CMV)carlier promoter and enhancer (pAd-CMV-VEGF165). The CHO cells was transfected pAdCMV-VEGF165. Biological activity was examened with Mile's test. Results:The CHO cells in culture after transfected pAd-CMV-VEGF165 demonstracted high expression,and Mile's test revealed that the VEGF protein which secreted from CHO cells can significantly enhanced vascular pemeability. Con-clusion:Our study established a good base for the further investigation of angiogenic gene therapy.
出处
《中国现代普通外科进展》
CAS
1999年第4期16-18,共3页
Chinese Journal of Current Advances in General Surgery
