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Evaluation of tyrosinase minigene co-injection as a marker for genetic manipulations in transgenic mice.

机译:评价酪氨酸酶小基因共注射作为转基因小鼠中基因操作的标志物。

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摘要

The utility of tyrosinase minigene co-injection was evaluated as a visual marker for the generation and breeding of transgenic mice. In an evaluation of 39 transgenic founder animals and 44 transgenic lines five phenotypic patterns of pigmentation were consistently observed, including albino, dark, light, mottled and himalayan. In these studies co-injection of the tyrosinase minigene along with the transgene of interest (TOI) resulted in genomic integration of the two transgenes in 95% of the F0 generation. Co-segregation of transgenes occurred in 94% of doubly transgenic mice in the F1 generation, without dissociation in subsequent generations. All pigmented phenotypes proved useful for distinguishing homozygous from heterozygous F2 animals via backcross trials, while the light, mottled and himalayan phenotypes proved useful in visually discriminating between homozygous and heterozygous F2 animals. In addition, the light, mottled and himalayan phenotypes proved useful in determining segregation patterns of transgenes in the progeny of crosses between separate transgenic lines. Moreover, there appears to be a correlation between intensity of pigmentation and degree of expression of the co-injected TOI. These studies confirm that tyrosinase co-injection is a useful adjunct in transgenic mouse studies and can serve to reduce routine genetic validation of transgenic lines.
机译:酪氨酸酶小基因共注射的效用被评估为转基因小鼠的产生和繁殖的视觉标记。在对39个转基因创始动物和44个转基因品系的评估中,始终观察到5种色素沉着的表型,包括白化病,深色,浅色,斑驳和喜马拉雅。在这些研究中,将酪氨酸酶小基因与目的转基因(TOI)一起注射会导致两个转基因在95%的F0代中进行基因组整合。转基因的共分离发生在F1代中94%的双转基因小鼠中,而在后代中没有解离。通过回交试验,所有有色表型都可用于区分纯合子和杂合子F2,而浅色,斑驳和喜马拉雅表型可用于在视觉上区分纯合子和杂合子F2。另外,轻,杂色和喜马拉雅表型被证明可用于确定分离的转基因系之间的杂交后代中的转基因分离模式。而且,在色素沉着强度和共注射TOI的表达程度之间似乎存在相关性。这些研究证实酪氨酸酶共注射在转基因小鼠研究中是有用的辅助手段,可用于减少转基因品系的常规遗传验证。

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