重组可能导致双荧光报告基因逆转录病毒转导中出现虚假结果。
Recombination can lead to spurious results in retroviral transduction with dually fluorescent reporter genes.
机构信息
Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, Missouri, USA.
出版信息
J Virol. 2013 Dec;87(24):13900-3. doi: 10.1128/JVI.02524-13. Epub 2013 Sep 25.
Abstract
Fluorescent proteins are routinely employed as reporters in retroviral vectors. Here, we demonstrate that transduction with retroviral vectors carrying a tandem-dimer Tomato (TdTom) reporter produces two distinct fluorescent cell populations following template jumping due to a single nucleotide polymorphism between the first and second Tomato genes. Template jumping also occurs between repeated sequences in the Tomato and green fluorescent protein (GFP) genes. Thus, proper interpretation of the fluorescence intensity of transduced cells requires caution.
摘要
荧光蛋白通常被用作逆转录病毒载体的报告基因。在这里，我们证明，携带串联二聚体番茄（TdTom）报告基因的逆转录病毒载体转导后，由于第一个和第二个番茄基因之间的单个核苷酸多态性，会因模板跳跃而产生两种不同的荧光细胞群体。番茄和绿色荧光蛋白（GFP）基因中的重复序列之间也会发生模板跳跃。因此，对转导细胞荧光强度的正确解释需要谨慎。
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