Transgene expression is associated with copy number and cytomegalovirus promoter methylation in transgenic pigs.
Transgene expression is associated with copy number and cytomegalovirus promoter methylation in transgenic pigs.
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DOI：
10.1371/journal.pone.0006679
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发表时间：
2009-08-18
期刊：
影响因子：
3.7
通讯作者：
Liu Z
中科院分区：
文献类型：
--
作者：
Kong Q;Wu M;Huan Y;Zhang L;Liu H;Bou G;Luo Y;Mu Y;Liu Z
转基因动物多年来一直被用来研究基因功能，产生重要的蛋白质，并为人类疾病的研究建立模型。然而，转基因动物的遗传和表达的不稳定性是一个主要的限制因素。拷贝数和启动子甲基化是已知的调节基因表达的因素，但还没有报道系统地研究它们对转基因表达的影响。在本研究中，我们用巨细胞病毒(CMV)驱动的体细胞核移植(SCNT)获得了两头表达绿色荧光蛋白(GFP)的转基因猪。采用实时荧光定量聚合酶链式反应和亚硫酸氢盐测序的方法检测转基因拷贝数和启动子甲基化水平。分析转基因猪个体发育、成纤维细胞、不同组织和后代中转基因表达与拷贝数和启动子甲基化的相关性。我们的结果表明，转基因猪的转基因表达与拷贝数和CMV启动子甲基化有关。
Transgenic animals have been used for years to study gene function, produce important proteins, and generate models for the study of human diseases. However, inheritance and expression instability of the transgene in transgenic animals is a major limitation. Copy number and promoter methylation are known to regulate gene expression, but no report has systematically examined their effect on transgene expression. In the study, we generated two transgenic pigs by somatic cell nuclear transfer (SCNT) that express green fluorescent protein (GFP) driven by cytomegalovirus (CMV). Absolute quantitative real-time PCR and bisulfite sequencing were performed to determine transgene copy number and promoter methylation level. The correlation of transgene expression with copy number and promoter methylation was analyzed in individual development, fibroblast cells, various tissues, and offspring of the transgenic pigs. Our results demonstrate that transgene expression is associated with copy number and CMV promoter methylation in transgenic pigs.
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