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目的 :检测含有变形链球菌葡糖基转移酶抗原基因 (gtfB)的重组质粒pcDNA3_gtfB能否在哺乳动物细胞COS_1中正确地转录和表达。方法 :采用脂质体介导法 ,将pcDNA3_gtfB转染哺乳动物细胞COS_1,采用RT_PCR法、免疫组化LSAB法、Western免疫印迹法检测重组质粒的转录水平和表达产物。结果 :pcDNA3_gtfB在转染COS_1细胞后具有转录和翻译活性 ,蛋白质表达产物分子量为 116～ 2 12kD ,且可与兔抗葡糖基转移酶抗体发生特异性结合。结论 :重组质粒pcDNA3_gtfB在哺乳动物细胞中能正确地转录和表达 ,为其作为基因疫苗进行动物实验提供了依据
OBJECTIVE: To detect whether the recombinant plasmid pcDNA3_gtfB containing the Streptococcus mutans glucosyltransferase gene (gtfB) can be correctly transcribed and expressed in mammalian cells COS_1. Methods: The pcDNA3_gtfB was transfected into mammalian cells COS_1 by liposome-mediated method. The RT-PCR, immunohistochemistry and Western blotting were used to detect the transcription level and expression of recombinant plasmid. Results: pcDNA3_gtfB had transcriptional and translational activities after transfection into COS_1 cells. The molecular weight of the expressed product was 116-212kD, and it could specifically bind with rabbit anti-glucosyltransferase antibody. CONCLUSION: The recombinant plasmid pcDNA3_gtfB can be correctly transcribed and expressed in mammalian cells, providing a basis for its use as a gene vaccine in animal experiments