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首页> 外文期刊>Fisheries and Aquatic Sciences >Comparative Analysis of Transgene Copy Numbers and Expression Characteristics across Multiple Transgenic Marine Medaka Oryzias dancena Strains carrying the β-Actin Promoter-Driven GFP Reporter
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Comparative Analysis of Transgene Copy Numbers and Expression Characteristics across Multiple Transgenic Marine Medaka Oryzias dancena Strains carrying the β-Actin Promoter-Driven GFP Reporter

机译：多个携带β-Actin启动子驱动的GFP报告基因的海洋Medaka Oryzias dancena菌株的转基因拷贝数和表达特征的比较分析

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Several transgenic marine medaka Oryzias dancena strains harboring a green fluorescent protein (GFP) reporter construct regulated by an endogenous -actin promoter were established and their expression characteristics in relation to transgene copy numbers were examined in 21 transgene genotypes. Most of the transgenic strains displayed transgene insertion patterns typical of microinjection-mediated introduction of foreign DNA into fish embryos, characterized by the random integration of multiple transgene copies (ranging from 1 - 282 copies per cell), often accompanied by the formation of concatemer(s), as assessed by genomic Southern blot hybridization analysis and qPCR. Transgenic strains showed ubiquitous and continued temporal and spatial expression patterns of the transgenic GFP during most of their life cycle, from the embryonic stage to adulthood, enabling assessment of the expression pattern of the endogenous -actin gene. However, a comparative evaluation of transgene copy numbers and expression levels showed that copy number-dependent expression, the stability of the ubiquitous distribution and expression efficiency per transgene copy varied among the transgenic strains. Fluorescence expression levels were positively correlated with absolute transgene copy numbers, whereas the expression efficiency per transgene copy was inversely related to the number of transgene integrant copies. Data from this study will guide the selection of potentially desirable transgenic strains with ubiquitous expression of a fluorescent transgene, not only in this marine medaka species but also in other related model fish species.
机译：建立了几种带有内源性肌动蛋白启动子调控的绿色荧光蛋白（GFP）报告基因构建体的海洋转基因稻高产稻瘟病菌菌株，并在21种转基因基因型中检测了它们与转基因拷贝数相关的表达特征。大多数转基因菌株表现出典型的转基因插入模式，这是显微注射介导的外源DNA进入鱼胚胎的典型特征，其特征是多个转基因拷贝（每个细胞1-282拷贝）的随机整合，通常伴随着串联体的形成（ s），通过基因组Southern blot杂交分析和qPCR评估。在从胚胎期到成年期的整个生命周期中，转基因菌株在转基因GFP的整个生命周期中都普遍存在并持续存在时间和空间表达模式，从而能够评估内源性肌动蛋白基因的表达模式。然而，对转基因拷贝数和表达水平的比较评估显示，转基因菌株之间拷贝数依赖性表达，遍在分布的稳定性和每个转基因拷贝的表达效率是不同的。荧光表达水平与绝对转基因拷贝数呈正相关，而每个转基因拷贝的表达效率与转基因整合体拷贝数成反比。这项研究的数据将指导选择具有潜在性的，普遍存在荧光转基因表达的转基因菌株的选择，不仅在该海洋中成虫中，而且在其他相关的模型鱼类中。

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