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| iNOS转基因在血管平滑肌细胞的表达 | |
| 引用本文： | 林京,张子力,黄晓玲,韩涛.iNOS转基因在血管平滑肌细胞的表达[J].福建医科大学学报,2006,40(4):327-329,333. |
| 作者姓名： | 林京 张子力 黄晓玲 韩涛 |
| 作者单位： | 1. 福建医科大学,生物化学与分子生物学系,福州,350004 2. 福建省立医院,心外科,福州,350001 3. 广东外语外贸大学,门诊部,广州,510420 |
| 基金项目： | 福建省自然科学基金;福建省科技厅科研项目;福建省卫生厅青年科研项目 |
| 摘 要： | 目的 研究逆转录病毒介导诱导型一氧化氮合酶（iNOS）基因在体外培养的大鼠主动脉平滑肌细胞（VSMC）中的表达。方法 通过脂质体介导的DNA转染法将PLXSNiNOS导入PA317细胞中；经G418筛选获得抗性克隆；用NIH3T3细胞测定病毒上清滴度；将病毒上清转染体外培养的VSMC；采用RT-PCR、Western blot检测VSMC内iNOS mRNA和蛋白的表达。结果 PLXSNiNOS转染体外培养的VSMC48h后，在VSMC内可检测到外源性iNOS mRNA和蛋白；而用PLXSN转染体外培养的VSMC48h后未能检测到外源性iNOS mRNA和蛋白表达。结论 逆转录病毒介导iN0s基因可高效转染体外培养的VSMC，并在细胞内表达iNOS蛋白。 |
| 关 键 词： | 一氧化氮合酶 基因转移 逆转录病毒科 肌 平滑 血管 细胞 培养的 |
| 文章编号： | 1672-4194（2006）04-0327-04 |
| 收稿时间： | 2006-03-03 |
| 修稿时间： | 2006-03-032006-06-25 |
The Expression of the Inducible Nitric Oxide Synthase Transgene in Cultured Vascular Smooth Muscle Cell by Transfection of Recombinant Retroviral Plasmid |
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| Lin Jing,Zhang Zili,Huang Xiaoling,Han Tao.The Expression of the Inducible Nitric Oxide Synthase Transgene in Cultured Vascular Smooth Muscle Cell by Transfection of Recombinant Retroviral Plasmid[J].Journal of Fujian Medical University,2006,40(4):327-329,333. | |
| Authors: | Lin Jing Zhang Zili Huang Xiaoling Han Tao |
| Institution: | 1. Division of Biochemistry and Molecular Biology, Fujian Medical University, Fuzhou 350004, China ;2. Department of Cardiovascular Surgery, Fujian Provincial Hospital, Fuzhou 350001, China ;3. Outpatient Department, Guangdong University of Foreign Studies, Guangzhou 510420, China |
| Abstract: | Objective To study the expression of inducible nitric oxide synthase(iNOS) gene mediated by retroviral vector in cultured aortic vascular smooth muscle cell(VSMC) of rat. Methods Recombinant retroviral vector PLXSN iNOS was transferred into the amphotropic packaging cell line PA317 by means of lipofectamine-mediated method.Anti-G418 positive clones were selected by G418.The viral titer was determined with the NIH3T3.Ex vitro VSMC were transfected by viral supernatant.The expression of the retroviral iNOS transgene was examined by RT-PCR and Western blot analyses. Results mRNA and protein of transferred iNOS gene were detected 48 hours after gene transfer within the transfected cells.Alone PLXSN-treated VSMC showed no evidence of iNOS mRNA and protein.Conclusion iNOS gene could be quickly and effectively transferred into cultured VSMC by retroviral vector. |
| Keywords: | nitric-oxide synthase gene transfer retroviridae muscle smooth vascular cells cultured |
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