BAC transgenic expression efficiency: bicistronic versus ATG-fusion strategies.
BAC transgenic expression efficiency: bicistronic versus ATG-fusion strategies.
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BAC 转基因表达效率：双顺反子与 ATG 融合策略。
DOI：
10.1002/dvg.20341
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Tsai,RobertYL
中科院分区：
文献类型：
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作者：
Lin,Tao;Yasumoto,Hiroaki;Tsai,RobertYL
基于细菌人工染色体（BAC）的转基因可以与靶基因双顺反子表达或与其翻译起始密码子融合。为了比较这两种方法的转基因表达效率，使用双顺反子（NSiGFP）或ATG-融合方法（NSmGFP）产生从核干细胞蛋白的基因组基因座表达绿色荧光蛋白（GFP）的小鼠。产生具有1、2和4个拷贝的NSiGFP转基因的三个品系和具有1个拷贝的NSmGFP转基因的两个品系。在三个NSiGFP系中，仅4拷贝系可以在其GFP蛋白表达水平上匹配NSmGFP系。GFP和核干细胞蛋白RNA转录物水平的分析排除了IRES无效性，并表明双顺反子信息的过早终止是NSiGFP小鼠中低GFP表达的原因。本工作为转基因表达水平至关重要时设计BAC转基因提供了重要信息。45：647-652，2007.© 2007 Wiley利斯公司
Bacterial artificial chromosome (BAC)‐based transgene can be expressed bicistronically with the target gene or fused to its translation start codon. To compare the transgene expression efficiencies of these two methods, mice were created that expressed green fluorescent protein (GFP) from the genomic locus of nucleostemin using the bicistronic (NSiGFP) or the ATG‐fusion approach (NSmGFP). Three lines with 1, 2, and 4 copies of the NSiGFP transgene, and two lines with 1 copy of the NSmGFP transgene were generated. Of the three NSiGFP lines, only the 4‐copy line can match the NSmGFP lines in their GFP protein expression levels. Analyses of the GFP and nucleostemin RNA transcript levels exclude IRES inefficiency and suggest premature termination of the bicistronic message as the cause for low GFP expression in the NSiGFP mice. This work provides important information for designing BAC transgenics when the transgene expression level is crucial. 45:647–652, 2007. © 2007 Wiley‐Liss, Inc.