Quantitative comparison of gene co-expression in a bicistronic vector harboring IRES or coding sequence of porcine teschovirus 2A peptide
Quantitative comparison of gene co-expression in a bicistronic vector harboring IRES or coding sequence of porcine teschovirus 2A peptide
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DOI：
10.1134/s1068162013040122
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发表时间：
2013-07-01
影响因子：
1
通讯作者：
Vinogradova, T. V.
中科院分区：
文献类型：
--
作者：
Kuzmich, A. I.;Vvedenskii, A. V.;Vinogradova, T. V.
在生物技术中，通常需要同时表达一个以上的靶基因。为此目的，正在积极开发编码几种蛋白质的多顺反子载体。最常见的是，市售载体利用脑心肌炎病毒（IRES EMCV）的各种类型的内部核糖体进入位点。然而，许多研究人员认为基于编码自切割2A肽的序列的双顺反子载体更有前途。在这项工作中，我们比较了在用双顺反子构建体转染的细胞中基因表达的效率，所述双顺反子构建体具有IRES EMCV或对应于猪捷申病毒-1 2A肽的P2 A核苷酸序列。通过测量编码由IRES或P2 A序列连接的RFP和EGFP蛋白的基因的共表达水平，在三种哺乳动物细胞系中确定基因表达效率。在用基于P2 A序列的遗传构建体转染的细胞中检测到更高水平的转基因表达。
In biotechnology, simultaneous expression of more than one target gene is often required. Multicistronic vectors encoding several proteins are being actively developed for this purpose. Most often, the commercially available vectors utilize various types of internal ribosomal entry site of the encephalomyocarditis virus (IRES EMCV). However, many researchers consider bicistronic vectors on the basis of sequences that encode self-cleaving 2A peptides more promising. In the work, we compare the efficiency of gene expression in cells transfected with bicistronic constructs bearing either IRES EMCV or the P2A nucleotide sequence corresponding to the porcine teschovirus-1 2A peptide. Efficiency of gene expression was determined in three mammalian cell lines by measurement of co-expression levels of genes coding for RFP and EGFP proteins linked by IRES or P2A sequence. A higher level of the transgene expression was detected in cells transfected with P2A sequence-based genetic constructs.