Analysis and expression of Pmlyzi3 from Penaeus monodon

¹ Shenzhen Key Laboratory of Marine Bioresource & Eco-environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, PR China
² Guangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, PR China
³ Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China

^* Corresponding author: dengxu@szu.edu.cn

Abstract

Lysozymes are crucial immune moleculars and play an important role in innate imunity. Here, a new lysozyme named Pmlyzi3 was found from the transcriptome data of Panaeus monodon. The Pmplyzi3 gene was 438bp in length, encoding a 146-residues peptide and the first 19 residues constituted a signal peptide. The mature peptide contained 10 cysteines and had 7 α-helixes in its N terminal. Moreover, it showed 88% identity with lysozyme-like protein from Penaeus vannamei. To express Pmlyzi3, pColdIV-SUMOPmlyzi3 plasmid was constructed by linked the Pmlyzi3 with SUMO tag, then transformed to Eschericha coli BL21 (DE3). By optimizing expression condition, SUMO-Pmlyzi3 was succeeded in expression in high level and purifing with Ni-NTA column. Following with SUMO protease excision, pure Pmlyzi3 was obtained by removing SUMO tag, which would be helped to study its function.