摘要
目的:将野生型和突变型(R778L)肝豆状核变性基因(ATP7B)转染入Me32aT22/2L细胞株中进行表达,观察野生型和突变型ATP7B的铜转运功能,为以后更深入的基因治疗奠定基础。方法:采用脂质体转染法分别将含野生型和突变型cDNA重组真核表达载体pRc/CMV-WD转染到Me32aT22/2L细胞株中,免疫荧光观察野生型和突变型ATP7B在细胞内的分布,以及铜孵育实验检查野生型和突变型ATP7B的铜转运功能。结果:在转染的Me32aT22/2L细胞株中可检测到ATP7B的表达,ATP7B位于细胞核的周围,在24和48小时后,细胞内铜/蛋白比值分别为335.33±49.86和477.38±30.95,而突变型ATP7B组铜/蛋白比值分别为606.14±45.72和901.84±53.18,两者比较具有统计学差异(P<0.05)。结论:野生型ATP7B具有铜转运功能,而突变型ATP7B则丧失了铜转运的功能。突变R778L基因是肝豆状核变性的致病基因。
Objective: Mutant-type and Wild-type ATP7B were transfected and expressed in skin fibroblasts Me32aT22/2L cell lines. Copper transport function of mutant-type and wild-type ATP7B were studied and made the basement for further gene therapy of hepatolenticular degeneration. Methods: pRc/CMV-WD containing cDNA of mutant-type or wild-type ATP7B was transfected into Me32aT22/2L cell using liposome transfection methods,respectively. Intracellular distribution of mutant-type and wild-type ATP7B were observed by im...
出处
《岭南急诊医学杂志》
2008年第6期403-404,429,共3页
Lingnan Journal of Emergency Medicine
基金
广东省医学科研基金(B2005038)资助
