热休克蛋白对HIV-1 Nef抗原在哺乳动物细胞中传递的影响

A. Milani, F. Rouhollah, Maryam Naseroleslami, A. Bolhassani
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引用次数: 3

摘要

48小时后,使用荧光显微镜和流式细胞术评估它们的转染效率。结果:成功地制备了pEGFP-nef、hsp27、hsp27-nef、hsp70和hsp70-nef结构体。用每种构建体转染细胞的结果显示,Nef-GFP、Hsp27-GFP、HSP7-Nef-GFP、Hsp70-GFP、Hsp70-Nef-GFP+Hsp70GFP+GFP和GFP的表达百分比为53.1% 0.2,64.22 0.8,57.1 0.7,68.8 1.0,61.7 0.7,77.4 1.5和81.8 1.8。结论:Hsp70对HEK-293T细胞Nef-GFP表达的促进作用大于Hsp27。此外,与pEGFP-hsp70-nef相比,pEGFP-nef和pEGFP-hsp70的递送增加了细胞中GFP群体的比率。这些构建体将在不久的将来用于在动物模型中开发HIV-1 DNA疫苗。
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The Effects of Heat Shock Proteins on Delivery of HIV-1 Nef Antigen in Mammalian Cells
. After 48 hours, their transfection efficiency was evaluated using fluorescent microscopy and flow cytometry. Results: The pEGFP-nef , - hsp27, -hsp27-nef, -hsp70 and - hsp70 - nef constructs were successfully prepared in large scale and high purity. The results of cell transfection with each construct showed that the percentages of Nef-GFP, Hsp27-GFP, Hsp27-Nef-GFP, Hsp70-GFP, Hsp70-Nef-GFP, Hsp70-GFP + Nef-GFP and GFP expression were 53.1  0.2, 64.22  0.8, 57.1  0.7, 68.8  1.0, 61.7  0.7, 77.4  1.5 and 81.8  1.8, respectively. Conclusion: These data showed that the potency of Hsp70 was more than Hsp27 for increasing Nef-GFP expression in HEK-293T cells. Moreover, the delivery of pEGFP-nef along with pEGFP-hsp70 increased the rate of GFP population in the cells as compared to that in pEGFP-hsp70-nef. These constructs will be used for development of HIV-1 DNA vaccine in animal model in near Future.
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