CAJ | 학술논문

背景：器官移植后的免疫排斥反应或免疫抑制药物严重不良反应使患者得不到有效治疗、治疗效果不佳。基于此背景下，结合最新免疫调节研究结果，试图寻找到一种有效的生物免疫抑制方法。 　　目的：构建过表达吲哚胺2，3-双加氧酶(indoleamine 2，3-dioxygenase，IDO)慢病毒载体。 　　方法：①将已构建成功的IDO基因插入慢病毒包装质粒GV308中，构建GV308-IDO重组慢病毒包装质粒。②用慢病毒包装辅助元件Psi、 cPPT、3FLAG、TetR、IRES、WRPE、TetIIP、Ubiquitin Promoter、SV40 origin及HIV的基本元件5’LTR 和3’LTR，共培养法转染80%融合的293T细胞。 　　结果与结论：Western blot检测经10 g/L 琼脂糖凝胶电泳可见在 Mr 48000处有一目的片段，该值与IDO蛋白大小一致。RT-PCR检测可见293T细胞中有IDO基因表达。说明IDO融合基因已经成功重组于慢病毒包装质粒内。
배경：기관이식후적면역배척반응혹면역억제약물엄중불량반응사환자득불도유효치료、치료효과불가。기우차배경하，결합최신면역조절연구결과，시도심조도일충유효적생물면역억제방법。 　　목적：구건과표체신타알2，3-쌍가양매(indoleamine 2，3-dioxygenase，IDO)만병독재체。 　　방법：①장이구건성공적IDO기인삽입만병독포장질립GV308중，구건GV308-IDO중조만병독포장질립。②용만병독포장보조원건Psi、 cPPT、3FLAG、TetR、IRES、WRPE、TetIIP、Ubiquitin Promoter、SV40 origin급HIV적기본원건5’LTR 화3’LTR，공배양법전염80%융합적293T세포。 　　결과여결론：Western blot검측경10 g/L 경지당응효전영가견재 Mr 48000처유일목적편단，해치여IDO단백대소일치。RT-PCR검측가견293T세포중유IDO기인표체。설명IDO융합기인이경성공중조우만병독포장질립내。
BACKGROUND:Immune rejections after organ transplantation or serious adverse reactions due to immunosuppressive drugs show a lack of effective treatments and poor therapeutic outcomes. Therefore, we try to find an effective immune suprresion method in combination of the latest immunomodulatory achievements. OBJECTIVE:To construct a lentiviral vector overexpressing indoleamine 2,3-dioxygenase (IDO). METHODS:(1) The IDO gene that was successful y contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cel s with 80%confluence were co-cultured with 5'LTR and 3'LTR, basic elements of lentiviral packaging auxiliary components, including Psi, cPPT, 3FLAG, TetR, IRES, WRPE, TetIIP, Ubiquitin Promoter, SV40 origin and HIV. RESULTS AND CONCLUSION:Western blot assay showed that in 10 g/L agarose gel electrophoresis, there was a target fragment at Mr 48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cel s. These findings suggest that IDO fusion gene has been successful y reorganized in the lentiviral packaging plasmids.