Constructing a lentiviral vector overexpressing indoleamine 2,3-dioxygenase

doi:10.3969/j.issn.2095-4344.2015.36.022

Abstract

Abstract:

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 successfully contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cells 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 M_r48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cells. These findings suggest that IDO fusion gene has been successfully reorganized in the lentiviral packaging plasmids.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Gene Fusion, Indoleamine-Pyrrole 2,3,-Dioxygenase, Lentivirus Infections

CLC Number:

R394.2

He Ji-gang, Li Hong-rong, Gui Long-sheng, Li Yong-wu, Yan Dan. Constructing a lentiviral vector overexpressing indoleamine 2,3-dioxygenase [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(36): 5859-5864.

Figures/Tables 4

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