摘要
目的:建立基因枪子弹制备及转染体外培养COS-7细胞系的方法。方法:以亚精氨、氯化钙沉淀法制备子弹(DNA+金颗粒),利用原子力显微镜观察子弹制备情况;采用基因枪方法分别将真核表达质粒pVax-Dsred-IRES-EGFP转染对照组和实验组COS-7细胞,转染后24h,利用激光扫描共聚焦显微镜观察细胞中红、绿荧光蛋白的表达。结果:制备了基因枪子弹,DNA紧密包裹在金颗粒周围;基因枪介导的pVax-Dsred-IRES-EGFP被转染入体外培养的COS-7细胞,转染后24h可检测到红、绿荧光,而对照组则没有荧光蛋白的表达。结论:国产新芝SJ-500型基因枪能够有效介导外源基因转移,基因枪转染的COS-7细胞能够有效表达报告基因。
Objective： To establish a preparation method of cartridge for gene gun and to observe the gene gun-mediated gene transfectiono to COS-7 cell lines in vitro. Methods： The cartridge（microcarrier＋DNA） was prepared by precipitation method of spermidine and calcium chloride. Using atomic force microscope to observe the detail of the cartridge. The recombinant eukaryotie expression vector pVax-Dsred-IRES-EGFP was tansfected into COS-7 cell lines by gene gun. The. expression of report gene was observed through laser scan confocal microscopy. Results： The cartridge of gene gun was successfully prepared, DNA-coated gold mierocarriers was achieved. Red and green fluorescent protein could be detected in the transfected COS-7 cell lines after 24 hours, when no fluorescent protein was detected in control group. Conclusion： SJ-500 gene gun-mediated gene transfer is effective and practicable. The report gene can be expressed through gene gun mediated transfection to cell lines in vitro.
出处
《生物技术通讯》
CAS
2008年第5期681-684,共4页
Letters in Biotechnology
基金
国家自然科学基金(30772002)
国家高技术研究发展计划(2006AA02A237
2007AA02Z451)
关键词
基因枪
细胞转染
真核表达载体
荧光蛋白
COS-7细胞系
gene gun
cell transfection
eukaryotic expression vector
fluorescent protein
COS-7 cell lines
