GJB6基因突变体过表达慢病毒载体的构建及其在HaCaT细胞中的表达

Construction of the over-expression lentivirus vector of the GJB6 gene and the expression in HaCaT cells

目的;明确GJB6目的基因蛋白Cx30在GJB6 3种突变体A88V、G11R和V37E在HaCaT细胞中的表达。方法:构建GJB6基因突变体的慢病毒载体,并转染至HaCaT,应用荧光倒置显微镜检测其表达。结果:突变体中G11R和A88V经过多次转染实验,能够明确观察到HaCaT细胞细胞膜上有Cx30蛋白的荧光表达;而V37E突变型荧光表达极弱或无。结论:3种突变体的Cx30表达量不同,初步推测突变体编码的蛋白质或不能定位于角质形成细胞膜或不能形成功能性的Cx30,从而影响了HaCaT细胞正常的增殖和分化。

Objective： To determine the expression of connexin-30 （Cx30） in three mutations （A88V, GllR and V37E） of GJB6 gene in HaCaT cells. Methods： The lentivirus vector of the gene GJB6 was constructed and transinfected to HaCaT cells. The expressions of Cx30 in HaCaT cells were detected by fluorescent inverted microscope. Results： The expression of Cx30 of A88V and G11R in HaCaT cells was significant and that of V37E was weak. Conclusion： The effects of the three mutations of GJB6 gene on the proliferation and differentiation of HaCaT may relate to the functional Cx30 and positioning on membranes of cells.