G418处理核供体细胞对猪克隆胚胎体外发育效率的影响
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863计划项目(2011AA100304);广东省自然科学基金(2014A030313464)


Effects of G418 treatment of donor cells on the in vitro developmental efficiency of cloned porcine embryos
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    【目的】探索G418处理供体细胞对其核移植胚胎发育效率的影响。【方法】利用G418单独处理猪成体成纤维细胞6 d后,收集细胞,利用荧光定量PCR的方法检测处理前后细胞中抗氧化应激、细胞凋亡相关基因的表达水平,运用亚硫酸盐结合测序法分别检测基因组重复序列LINE-1、微卫星的DNA甲基化状态,以及其核移植胚胎体外发育的能力。【结果】经不同质量浓度G418处理的供体细胞的抗氧化应激酶相关基因及细胞凋亡基因表达发生显著变化(P<0.05),但其DNA甲基化水平没有发生改变(P>0.05);经G418处理的供体细胞的核移植胚胎的体外发育效率显著低于对照组(P<0.05)。【结论】G418处理供体细胞可能对其克隆胚胎体外发育效率有抑制作用。

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    【Objective】To explore the influence of G418 treatment of donor cells on the developmental efficiency of somatic cell nuclear transfer (SCNT) embryos.【Method】Porcine adult fibroblasts were collected after six days of G418 treatment. Expression levels of antioxidation and apoptosis-related genes were detected by quantitative RT-PCR before and after G418 treatment. The DNA methylation levels of LINE-1 repetitive sequences and microsatellites were analyzed using bisulfites sequencing analysis, and the in vitrodevelopmental rates of SCNT embryos were investigated.【Result】G418 treatment of donor cells caused significant (P<0.05) changes in expression levels of genes related to antioxidation and apoptosis, but did not significantly(P>0.05) change DNA methylation levels. SCNT embryos cloned from G418-treated donor cells exhibited a significantly (P<0.05) lower in vitro developmental rates compared with the control group.【Conclusion】G418 treatment of donor cell may inhibit the development of SCNT embryo.

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李嘉琪,李紫聪,吴珍芳,刘德武. G418处理核供体细胞对猪克隆胚胎体外发育效率的影响[J].华南农业大学学报,2016,37(5):13-18

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  • 收稿日期:2016-01-09
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  • 在线发布日期: 2016-07-25