Expression and Structural Prediction of Adenovirus Hex Protein

According to the sequence of CAdV-2 Hex (GenBank accession number AC000020.1), the primers were designed to amplify an 897-bp fragment (F: 5′-1654-TATGAATTCGACCTCCGAGTGGATGGG-1671-3′, R: 5′-2533-GAGCTCGAGGGAGTTAGAGTACAGCAG—2550-3′). The fragment was cloned into a pET30a (+) vector (Tiangen, Beijing, China) to construct the expression plasmids. The expression plasmid was transformed into E. coli BL₂₁ competent cells (Tiangen); the truncated Hex protein was produced via isopropyl-β-D-thiogalactopyranoside (IPTG, GE Healthcare, Chicago, IL, USA) induction using pET30a- Hex E. coli at 37 °C. The truncated Hex protein was detected by SDS-PAGE and Western blotting (WB) analysis by anti-His mAb. Immunoreactive bands were verified by an enhanced chemiluminescence system (ECL; PerkinElmer Life Sciences, Fremont, CA, USA) [15 (link)]. The three-dimensional (3D) structure of the expressed protein was predicted with PyMOL software based on data from the SWISS-MODEL online server.