Sequence characterization and expression of Trichinella spiralis 5'-nucleotidase gene
LIAO Cheng-shui1,3, WANG Xiao-li2, DU Fu-yu1, YU Chuan1, YU Zu-hua1, ZHANG Chun-jie1, LI Yin-ju1, WU Ting-cai1, LIU Ming-yuan3, CHENG Xiang-chao1
1. College of Animal Science and Technology /Luoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control, Henan University of Science and Technology, Luoyang 471023, China; 2. Medical College, Henan University of Science and Technology, Luoyang 471023, China; 3. Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, Jilin University, Changchun 130062, China
Abstract：We aimed to study the sequence structure and cloning and expression of 5'-nucleotidase gene from Trichinella spiralis (T. spiralis). The sequence of 5'-nucleotidase gene was obtained from T. spiralis, analyzed systematically by bioinformatics method, and expressed in prokaryotic expression system and purified. Sequencing analysis showed that the size of 5'-nucleotidase gene was 1 653 bp. The gene encoded a protein of 550 amino acids. The protein was composed of 119 amino acid residues and the molecular formula was C2800H4358N742O803S23. 5'-nucleotidase was a stable hydrophilic protein with a predicted molecular mass (Mr) of 62 kD and theoretical pI of 6.13. The protein has signal peptides and a transmembrane region. A total of 3 N-glycosylation sites, 2 O-glycosylation sites, 20 phosphorylation sites, 28 B cells linear binding sites, and 13 T cell binding sites were predicted in 5'-nucleotidase. In the secondary structure, α-helix accounted for 43.27% (238), extension chain accounted for 22.73% (125), β-sheet accounted for 7.82% accounted for 26.18% (144). SDS-PAGE analysis showed that the 5'-nucleotidase gene could be successfully expressed in prokaryotic expression system. The soluble expression products with about 74 kD were purified by Ni-NTA affinity chromatography. In this study, the 5'-nucleotidase gene from T. spiralis was successfully cloned and analyzed and highly expressed in E. coli. The results laid the foundation for the further study on the role of 5'-nucleotidase of development infection in T. spiralis.
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2018, Vol. 34 
