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目的:建立基因枪子的弹制备及将不同质粒DNA转染体外培养MCF-7细胞系的方法。方法:以亚精氨-氯化钙沉淀法制备子弹,采用基因枪方法分别将真核表达质粒pEG-FP,Pmcherry转染对照组和实验组MCF-7细胞,转染24h后,利用荧光显微镜观察细胞中红、绿荧光蛋白的表达情况。结果:制备了基因枪子弹,基因枪介导的pEGFP、Pmcherry能够分别和共同转染入体外培养的MCF-7细胞中,转染后24h可检测到红、绿色荧光,而对照组则没有荧光蛋白的表达。结论:基因枪能够有效介导外源基因转移并能够实现两种基因的共同转移,基因枪转染的MCF-7细胞能够有效表达报告基因。
OBJECTIVE: To establish the bullet preparation of gene gun and the method of transfecting different plasmid DNA into MCF-7 cell line in vitro. Methods: The bullets were prepared by sub-ammonia-calcium chloride precipitation method. The eukaryotic expression plasmids pEG-FP and Pmcherry were transfected into control group and experimental group MCF-7 cells respectively by gene gun method. After 24h transfection, Observe the expression of red and green fluorescent protein in cells. Results: Gene gun bullets were prepared. Gene gun-mediated pEGFP and Pmcherry were transfected into MCF-7 cells separately and co-transfected. Fluorescence red and green fluorescence were detected 24h after transfection, but no fluorescence Protein expression. CONCLUSION: Gene gun can effectively mediate the exogenous gene transfer and can co-transfer the two genes. The gene gun transfected MCF-7 cells can effectively express the reporter gene.