Kim Sujeong, Lee Karim, Kim Mi-Dong, Kang Seock, Joo Chang Wan, Kim Jong-Mook, Kim Seon Hee, Yu Seung Shin, Kim Sunyoung
ViroMed Co., Ltd., 1510-8 Bongcheon7-Dong, Gwanak-Gu, Seoul 151-818, Republic of Korea.
Biochem Biophys Res Commun. 2006 May 19;343(4):1017-22. doi: 10.1016/j.bbrc.2006.03.035. Epub 2006 Mar 20.
The long terminal repeat (LTR) of retrovirus contains the nucleotide sequences that control gene expression. Although several different LTRs have been used in the context of retroviral vector, the activity of the various LTRs has not yet been systematically compared for their level of gene expression. We evaluated the effect of four different LTRs on gene expression using luciferase, stem cell factor, and enhanced green fluorescence protein as reporter genes. LTRs tested in this study were derived from Moloney murine leukemia virus, myeloproliferative sarcoma virus, murine stem cell virus, and spleen focus-forming virus. It was found that the level of gene expression is affected by not only LTRs but also the transgenes and the cell types in which gene expression occurs. Furthermore, the presence of other nucleotide sequences such as the internal ribosome entry site (IRES)-neo cassette could also significantly affect gene expression. Our results suggested that the LTR should be chosen carefully, more or less on an empirical basis.
逆转录病毒的长末端重复序列(LTR)包含控制基因表达的核苷酸序列。尽管在逆转录病毒载体的背景下已经使用了几种不同的LTR,但尚未对各种LTR的基因表达水平进行系统比较。我们使用荧光素酶、干细胞因子和增强型绿色荧光蛋白作为报告基因,评估了四种不同LTR对基因表达的影响。本研究中测试的LTR分别来源于莫洛尼鼠白血病病毒、骨髓增殖性肉瘤病毒、小鼠干细胞病毒和脾脏灶形成病毒。研究发现,基因表达水平不仅受LTR的影响,还受转基因以及发生基因表达的细胞类型的影响。此外,其他核苷酸序列如内部核糖体进入位点(IRES)-新霉素盒的存在也会显著影响基因表达。我们的结果表明,应或多或少基于经验谨慎选择LTR。