核定位信号肽和转运肽对斑马鱼转基因效率的影响

为了探讨核定位信号肽S413-PV和转运肽Tp10这2种融合肽对转基因效率的影响,将S413-PV和Tp10与线性化质粒分别以质量比50∶1、100∶1、200∶1混合制成转染液,注射斑马鱼单细胞期胚胎,分别检测转染24 h、120 h后胚胎的阳性率和成活率。结果表明:与对照组相比,转染24 h后,S413-PV组胚胎阳性率极显著上升,胚胎成活率均下降,但差异不显著;Tp10组阳性率均呈上升趋势,200∶1组差异极显著,其他组差异不显著,胚胎成活率均显著下降;转染120 h后,S413-PV组胚胎阳性率均极显著上升,胚胎成活率均呈下降趋势,100∶1、200∶1组差异极显著;Tp10组胚胎阳性率上升,200∶1组差异显著,胚胎成活率均极显著下降。以上结果表明,S413-PV可显著提高斑马鱼转基因效率,对胚胎毒性较小;Tp10对转基因效率影响不显著,且对胚胎毒性较强。

Effect of Nuclear Localization Signal Peptides and Transit Peptides on Gene Transfer Efficiency in Zebrafish

In order to study effect of nuclear localization signal S413-PV and transport peptides Tp10 on gene transfer efficiency,the synthetic NLS peptide S413-PV or transport peptide Tp10 was mixed with li-nearized plasmid harbouring green fluorescent protein(GFP) expressing cassete at the mass ratio of 50∶1,100∶1,and 200∶1,respectively.Then the mixture was injected into 1-cell stage fertilized eggs of zebrafish.GFP positive efficiency and the survival rate of embryos were detected at 24 hours post fertilization(hpf) and 120 hpf stages.The results showed that the GFP positive rate of embryos in S413-PV groups were significantly higher than that in the control group both at 24 and 120 hpf stages.There were significant differences between the different ratio.The GFP positive rate of embryos was highest at the ratio of 200∶1.The GFP positive rates of embryos in Tp10 groups were also higher than that in the control at 24 and 120 hpf.And the GFP positive rate of embryos at the ratio of 200∶1 was significantly higher than that in other groups at 24 hpf.The survival rate of embryos either in S413-PV or Tp10 was lower than control.The rate decreased with the peptides ratio increased.The survival rate of embryos in S413-PV groups at the ratio of 100∶1 and 200∶1 were significantly lower than that in other groups at 120 hpf.The survial rate of embryos in Tp10 groups were significantly lower than that in control group both at 24 and 120 hpf stages.These data suggested that S413-PV could significantly improve the transgenic efficiency in zebrafish and had a little effect on the survival rate of embryos.While Tp10 had no significant effect on transgenic efficiency,and had higher toxicity to embryos.