论文部分内容阅读
目的 :建立血管生成素 (angiogenin ,ANG)基因的真核表达系统。方法 :采用化学合成拼接法 ,获得人ANG全长cDNA并测序 ,构建重组荧光真核细胞表达质粒 pEGFP ANG ,然后经脂质体介导转染人脐静脉内皮细胞 (HUVEC)。应用荧光显微镜观测及免疫组化染色 ,对转染细胞内 pEGFP ANG的表达进行鉴定。结果 :测序表明 ,编码的氨基酸序列与人ANG完全一致。荧光显微镜下可见转染的HUVEC内发出强绿色荧光。免疫组化检测显示 ,转染的HUVEC中有棕色颗粒。结论 :获得了人ANG的全长编码基因。构建的重组体pEGFP ANG转染HUVEC后 ,可表达人ANG蛋白
Objective: To establish an eukaryotic expression system of angiogenin (ANG) gene. Methods: Full-length human ANG cDNA was obtained by chemical synthetic splicing. The recombinant plasmid pEGFP ANG was constructed and transfected into human umbilical vein endothelial cells (HUVEC) by liposome. The expression of pEGFP ANG in transfected cells was identified by fluorescence microscopy and immunohistochemical staining. Results: Sequencing showed that the encoded amino acid sequence was completely consistent with human ANG. Fluorescent microscopy revealed strong green fluorescence in transfected HUVECs. Immunohistochemistry showed brown particles in transfected HUVECs. Conclusion: The full-length coding gene of human ANG was obtained. The constructed recombinant pEGFP ANG transfected HUVEC, human ANG protein can be expressed