A ''humanized'' green fluorescent protein cDNA adapted for high-level expression in mammalian cells
A ''humanized'' green fluorescent protein cDNA adapted for high-level expression in mammalian cells
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DOI：
10.1128/jvi.70.7.4646-4654.1996
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发表时间：
1996-07-01
影响因子：
5.4
通讯作者：
Muzyczka, N
中科院分区：
文献类型：
--
作者：
Zolotukhin, S;Potter, M;Muzyczka, N
我们构建了绿色荧光蛋白（h），一个合成版本的水母维多利亚水母绿色荧光蛋白（gfp）cDNA，适用于高水平表达的哺乳动物细胞，特别是那些人的起源。为了改变gfp 10编码序列中的密码子使用，在88个密码子中进行了总共92个碱基的替换，使得其更适合于在哺乳动物细胞中表达。我们还描述了一系列用于将基因递送和表达到哺乳动物细胞中的通用重组腺相关病毒和腺病毒载体，并且使用这些载体，证明GFP（H）基因在人细胞系293中以及在豚鼠眼的神经感觉细胞中的体内有效转导和表达。用重组腺相关病毒-GFP（H）感染的细胞可以很容易地通过荧光激活细胞分选术进行分选，这表明新设计的gfp（h）基因可以作为报告基因广泛地用于许多基因递送技术，包括人类基因治疗。
We constructed gfp(h), a synthetic version of the jellyfish Aequorea victoria green fluorescent protein (gfp) cDNA that is adapted for high-level expression in mammalian cells, especially those of human origin. A total of 92 base substitutions were made in 88 codons in order to change the codon usage within the gfp10 coding sequence so that it was more appropriate for expression in mammalian cells, We also describe a series of versatile recombinant adeno-associated virus and adenovirus vectors for delivery and expression of genes into mammalian cells and, using these vectors, demonstrate the efficient transduction and expression of the gfp(h) gene in the human cell line 293 and also in vivo, within neurosensory cells of guinea pig eye. Cells infected with recombinant adeno-associated virus-GFP(H) can be readily sorted by fluorescence-activated cell sorting, suggesting that the newly designed gfp(h) gene could he widely used as a reporter in many gene delivery technologies, including human gene therapy.