Construction of a recombinant adenovirus overexpressing rat TIPE2 gene / 中国组织工程研究

ABSTRACT

BACKGROUND:Tumor necrosis factor-α-induced protein-8 like-2 (TIPE2), an anti-inflammatory protein, through the T cel receptor (TCR) and TOLL-like receptor signaling pathway, implements negative regulation of adaptive immunity and innate immunity, and thus effectively maintains the stable internal environment of the body. OBJECTIVE:To construct a recombinant adenovirus that can overexpress rat TIPE2 gene. METHODS:TIPE2 cDNA target gene was amplified from rat’s lymphocytes using RT-PCR, cloned into shuttle plasmid pShuttle-clontech, and then subcloned into artificial adenovirus vector AdC68. Hereafter, HEK 293 cel s were transfected to generate a recombinant adenovirus. HEK293A cel s were infected using this recombinant adenovirus, and then TIPE2 gene level was tested by western blot method. RESULTS AND CONCLUSION:Based on results of PCR, digestion identification and sequencing, the obtained cDNA was the coding sequence region of TIPE2. Western blot findings showed that the recombinant adenovirus could overexpress TIPE2 gene. These findings indicate that the recombinant adenovirus is constructed successful y and can express TIPE2 gene stably.