Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL...Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL3-CMV, pGL3MyoGpro, pGL3-CMV-MyoGpro, and pGL3-Basic which contained CMV promoter, MyoG promoter, CMV-MyoG synthesis promoter, and a promoterless negative control, respectively. Then the four vectors and internal control Renilla luciferase report gene vector phRL-TK were transfected into bovine skeletal muscle satellite cells, mouse C2C12 cells and bovine fetal fibroblast cells to detect the promoter activity with dual luciferase report system. The results showed that CMV enhancer could significantly improve the transcription activity of bovine MyoG gene promoter in muscle satellite cells and mouse C2C12 cells, and it had certain specificity. This study provided experimental materials for increasing the high expression of exogenous gene in bovine muscle cells, and also laid the molecular theoretical basis for obtaining the high specific promoter of bovine muscle and the transgenic beef cattle.展开更多
目的 增强E .colicd/HSV 1tk自杀基因的细胞毒性 ,实现阳离子脂质体介导pE CEA cd tk/5 FC +GCV体系靶向杀伤CEA阳性肺癌。方法 PCR法分别扩增出CMV增强子 ,CEA启动子 ,cd tk ,构建真核表达载体 pE CEA cd tk ;MTT法检测pE CEA cd tk/...目的 增强E .colicd/HSV 1tk自杀基因的细胞毒性 ,实现阳离子脂质体介导pE CEA cd tk/5 FC +GCV体系靶向杀伤CEA阳性肺癌。方法 PCR法分别扩增出CMV增强子 ,CEA启动子 ,cd tk ,构建真核表达载体 pE CEA cd tk ;MTT法检测pE CEA cd tk/5 FC +GCV体系的体外细胞毒性。体内实验采用肺腺癌细胞SPC A 1裸鼠皮下移植瘤模型。通过肿瘤局部或鼠尾静脉注射脂质体 /pE CEA cd tk复合物 ,腹腔注射GCV +5 FC前体药物进行治疗。结果 阳离子脂质体介导pE CEA cd tk/5 FC +GCV体系体外可靶向杀伤CEA阳性肺癌细胞 ,这种杀伤作用存在显著的细胞差异 ;体内可抑制小鼠皮下肺肿瘤结节的生长 ,荷瘤鼠存活期延长。结论 阳离子脂质体介导 pE CEA cd tk/5 FC +GCV体系体内外对CEA阳性肺癌均具有较强的杀伤作用。展开更多
基金Supported by the Major Special Projects of New Product Training of Transgenic Organisms(zx080072008-2008)
文摘Connected a segment of CMV enhancer to the front of MyoG gene promoter and then constructed the corresponding dual luciferase expression vector pGL3-CMV-MyoGpro. We set four eukaryotic expression vectors including pGL3-CMV, pGL3MyoGpro, pGL3-CMV-MyoGpro, and pGL3-Basic which contained CMV promoter, MyoG promoter, CMV-MyoG synthesis promoter, and a promoterless negative control, respectively. Then the four vectors and internal control Renilla luciferase report gene vector phRL-TK were transfected into bovine skeletal muscle satellite cells, mouse C2C12 cells and bovine fetal fibroblast cells to detect the promoter activity with dual luciferase report system. The results showed that CMV enhancer could significantly improve the transcription activity of bovine MyoG gene promoter in muscle satellite cells and mouse C2C12 cells, and it had certain specificity. This study provided experimental materials for increasing the high expression of exogenous gene in bovine muscle cells, and also laid the molecular theoretical basis for obtaining the high specific promoter of bovine muscle and the transgenic beef cattle.
文摘目的 增强E .colicd/HSV 1tk自杀基因的细胞毒性 ,实现阳离子脂质体介导pE CEA cd tk/5 FC +GCV体系靶向杀伤CEA阳性肺癌。方法 PCR法分别扩增出CMV增强子 ,CEA启动子 ,cd tk ,构建真核表达载体 pE CEA cd tk ;MTT法检测pE CEA cd tk/5 FC +GCV体系的体外细胞毒性。体内实验采用肺腺癌细胞SPC A 1裸鼠皮下移植瘤模型。通过肿瘤局部或鼠尾静脉注射脂质体 /pE CEA cd tk复合物 ,腹腔注射GCV +5 FC前体药物进行治疗。结果 阳离子脂质体介导pE CEA cd tk/5 FC +GCV体系体外可靶向杀伤CEA阳性肺癌细胞 ,这种杀伤作用存在显著的细胞差异 ;体内可抑制小鼠皮下肺肿瘤结节的生长 ,荷瘤鼠存活期延长。结论 阳离子脂质体介导 pE CEA cd tk/5 FC +GCV体系体内外对CEA阳性肺癌均具有较强的杀伤作用。
