Author: Chen, Shilong; Wang, Long; Chen, Jieying; Zhang, Lanlan; Wang, Song; Goraya, Mohsan U.; Chi, Xiaojuan; Na, Yang; Shao, Wenhan; Yang, Zhou; Zeng, Xiancheng; Chen, Shaoying; Chen, Ji-Long
Title: Avian Interferon-Inducible Transmembrane Protein Family Effectively Restricts Avian Tembusu Virus Infection Document date: 2017_4_20
ID: 0pjg25kn_15
Snippet: Full-length cDNA encoding chicken or duck species IFITM1 and IFITM3 was subcloned into pFLAG-CMV-5a vector with a Flag tag in the COOH terminus to create DNA constructs chIFITM1, chIFITM3, duck IFITM1 (dIFITM1), and dIFITM3, respectively. The specific primers with restriction enzyme sites are shown in Table 1 . DF-1 cells were seeded onto 6-well plates and cultured in DMEM with 10% FBS. When the cells reached 60-80% confluence, they were transfec.....
Document: Full-length cDNA encoding chicken or duck species IFITM1 and IFITM3 was subcloned into pFLAG-CMV-5a vector with a Flag tag in the COOH terminus to create DNA constructs chIFITM1, chIFITM3, duck IFITM1 (dIFITM1), and dIFITM3, respectively. The specific primers with restriction enzyme sites are shown in Table 1 . DF-1 cells were seeded onto 6-well plates and cultured in DMEM with 10% FBS. When the cells reached 60-80% confluence, they were transfected with 3 µg of plasmid DNA/well using Lipofectamine 3000 (Invitrogen). Twenty four hours later, transfected DF-1 cells were infected with ATMUV CJD05 and harvested at indicated times (24, 36, and 48 hpi) for further qRT-PCR or Western blotting analysis. Supernatants were collected for detection of viral titers by TCID 50 assay.
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