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首页> 外文期刊>Nucleic Acids Research >HIGH-TITER BICISTRONIC RETROVIRAL VECTORS EMPLOYING FOOT-AND-MOUTH DISEASE VIRUS INTERNAL RIBOSOME ENTRY SITE
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HIGH-TITER BICISTRONIC RETROVIRAL VECTORS EMPLOYING FOOT-AND-MOUTH DISEASE VIRUS INTERNAL RIBOSOME ENTRY SITE

机译：运用双足病病毒的高双歧性逆转录病毒载体内部核糖体进入位点

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摘要

Bicistronic retroviral vectors were constructed containing the foot-and-mouth disease virus (FMDV) internal ribosome entry site (IRES) followed by the coding region of beta-galactosidase (beta-gal) or therapeutic genes, with the selectable neomycin phosphotransferase gene under the control of the viral long terminal repeat (LTR) promoter, LNFX, a vector with a multiple cloning site 3' to foot-and-mouth disease virus IRES, was used to construct vectors encoding rat erythropoietin (EP), rat granulocyte colony-stimulating factor (G-CSF), human adenosine deaminase (ADA) and beta-gal. In transduced primary rat vascular smooth muscle cells the cytokines were expressed at high levels, similar to those obtained from vectors employing the viral LTR promoter, LNFZ, a vector encoding beta-gal, had a 10-fold increase in titer over that of LNPoZ, a comparable vector containing the poliovirus (Po) internal ribosome entry site, Primary canine vascular smooth muscle cells infected with LNFZ and LNPoZ expressed similar activities of beta-gal and neomycin phosphotransferase (NPT), Overall, these vectors had titers between 10(6) and 2 x 10(7) c.f.u./ml, indicating that foot-and-mouth disease virus IRES provides high-titer bicistronic vectors with high-level two gene expression.
机译：构建了双顺反子逆转录病毒载体，其中包含口蹄疫病毒（FMDV）内部核糖体进入位点（IRES），其后是β-半乳糖苷酶（β-gal）或治疗性基因的编码区，在该区下有新霉素磷酸转移酶基因病毒长末端重复序列（LTR）启动子的控制，LNFX，一种对口蹄疫病毒IRES具有3'多克隆位点的载体，被用于构建编码大鼠促红细胞生成素（EP），刺激大鼠粒细胞集落的载体因子（G-CSF），人腺苷脱氨酶（ADA）和β-gal。在转导的大鼠原代血管平滑肌细胞中，细胞因子高水平表达，类似于从采用病毒LTR启动子的载体获得的细胞因子，LNFZ（编码β-gal的载体）的滴度比LNPoZ的滴度增加了10倍，包含脊髓灰质炎病毒（Po）内部核糖体进入位点的可比载体，感染LNFZ和LNPoZ的犬科动物血管平滑肌细胞表达类似的β-gal和新霉素磷酸转移酶（NPT）活性，总体而言，这些载体的效价在10（6）之间和2 x 10（7）cfu / ml，表明口蹄疫病毒IRES提供了具有高水平两个基因表达的高滴度双顺反子载体。

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