Objective: To construct a HIV-1 gp120 transgenic mice (gp120 Tg) with vimentin (VIM) gene knockout.
Methods: Female HIV-1 gp120 Tg mice were mated to VIM heterozygote mice (F0). All the offspring mice were derived from these original founders so that both genotypes had the same mixed genetic background. The F1 mice were bred to generate of VIM+/+, VIM-/-, VIM+/+/gp120 Tg and VIM-/-/gp120 Tg mice. PCR was performed for genotyping of the mice, and the expressions of VIM and gp120 in the brain tissues were examined using immunoblotting.
Results: The results of PCR showed the presence of the target bands in VIM+/+, VIM-/-, VIM+/+/gp120 Tg and VIM-/-/gp120 Tg mice. In VIM-/-/gp120 Tg mice, gp120 expression was detected throughout the brain regions while no VIM expression was detected.
Conclusions: We generated gp120 transgenic mouse models with VIM gene knockout, which facilitate the exploration of the role of VIM in gp120-induced neurotoxicity.
目的: 构建波形蛋白(VIM)基因敲除和gp120转基因(gp120 Tg)小鼠模型。
方法: 将VIM基因敲除小鼠(VIM-/-)和gp120Tg小鼠杂交(F0代), 然后在产生的子代(F1)中, 取分组为VIM+/-/gp120Tg的小鼠一公一母杂交, 得到基因型分别为VIM+/+、VIM+/-、VIM-/-和VIM+/+/gp120 Tg、VIM+/-/gp120 Tg、VIM-/-/gp120 Tg的6组小鼠。其中VIM+/+、VIM-/-、VIM+/+/gp120Tg和VIM-/-/gp120这4组小鼠互为对照。用PCR法鉴定这上述6组小鼠的基因型, 免疫印迹法检测小鼠脑组织中VIM和gp120的表达情况。
结果: 4组基因型VIM+/+、VIM-/-、VIM+/+/gp120Tg和VIM-/-/gp120Tg小鼠的PCR结果符合预期, 免疫印迹结果显示VIM-/-/gp120Tg小鼠VIM无表达, gp120过表达(P < 0.001), 符合预期结果。
结论: 成功构建了VIM基因敲除gp120转基因小鼠模型, 为深入研究波形蛋白在gp120神经毒性作用中的作用机制提供了坚实的研究基础。
Keywords: HIV; gp120; neurotoxicity; transgenic mice; vimentin.