PMID- 31794842 OWN - NLM STAT- MEDLINE DCOM- 20200406 LR - 20200408 IS - 1095-9947 (Electronic) IS - 1050-4648 (Linking) VI - 96 DP - 2020 Jan TI - Molecular cloning, expression analyses and functional characterization of a goose-type lysozyme gene from Bostrychus sinensis (family: Eleotridae). PG - 41-52 LID - S1050-4648(19)31110-6 [pii] LID - 10.1016/j.fsi.2019.11.067 [doi] AB - In this study, we sequenced and characterized the goose-type lysozyme gene, termed as BsLysG, from the Chinese black sleeper (Bostrychus sinensis). The BsLysG encodes 196 amino acids and contains a soluble bacterial lytic transglycosylases domain, three catalytic residues (Glu(72), Asp(85) and Asp(102)) and the GLMQ motif (Gly(97), Leu(98), Met(99) and Gln(100)). No signal peptide was observed in the BsLysG protein. The genomic DNA of BsLysG contains five exons and four introns. The sequence analyses showed that the BsLysG exhibits high similarity with LysG from other fishes. Phylogenetic analyses showed that the BsLysG is clustered together with its counterparts from other teleost fishes. The Real-time PCR analyses showed that the BsLysG was found to be ubiquitously expressed in ten examined organs in Chinese black sleeper, with predominant expression in spleen, followed by head kidney and peripheral blood. Expression analyses showed that the BsLysG was significantly upregulated in vivo after either pathogen Vibrio parahemolyticus infection or poly (I:C) challenge in peripheral blood, head kidney, liver and spleen organs. The purified recombinant BsLysG (rBsLysG) has optimal activity at 35 degrees C and pH 5.5. The rBsLysG exhibited antimicrobial activity against two Gram-positive bacteria (Micrococcus lysodeikticus and Staphylococcus aureus) and two Gram-negative bacteria (Escherichia coli and V. parahemolyticus). The Scanning electron microscope (SEM) imaging analyses showed that the rBsLysG-treated V. parahemolyticus cells displayed morphological deformation. These results indicate that the BsLysG is involved in host immune defense against bacterial infection. CI - Copyright (c) 2019 Elsevier Ltd. All rights reserved. FAU - Wei, Ke AU - Wei K AD - National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan, 316004, China. FAU - Ding, Yuehan AU - Ding Y AD - National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan, 316004, China. FAU - Yin, Xiaolong AU - Yin X AD - Zhoushan Fisheries Research Institute, Zhoushan, Zhejiang, 316022, China. FAU - Zhang, Jianshe AU - Zhang J AD - National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan, 316004, China. Electronic address: zhangjianshe@zjou.edu.cn. FAU - Shen, Bin AU - Shen B AD - National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan, 316004, China. Electronic address: shenbin@zjou.edu.cn. LA - eng PT - Journal Article DEP - 20191130 PL - England TA - Fish Shellfish Immunol JT - Fish & shellfish immunology JID - 9505220 RN - 0 (Fish Proteins) RN - EC 3.2.1.17 (Muramidase) RN - O84C90HH2L (Poly I-C) SB - IM MH - Amino Acid Sequence MH - Animals MH - Base Sequence MH - Fish Diseases/*immunology MH - Fish Proteins/chemistry/genetics/immunology MH - Fishes/*genetics/*immunology MH - Gene Expression Profiling/veterinary MH - Gene Expression Regulation/*immunology MH - Gram-Negative Bacteria/physiology MH - Gram-Positive Bacteria/physiology MH - Immunity, Innate/*genetics MH - Muramidase/chemistry/*genetics/*immunology MH - Phylogeny MH - Poly I-C/pharmacology MH - Sequence Alignment/veterinary OTO - NOTNLM OT - Chinese black sleeper OT - Gene expression OT - Goose-type lysozyme OT - Innate immunity OT - Lytic activity EDAT- 2019/12/04 06:00 MHDA- 2020/04/09 06:00 CRDT- 2019/12/04 06:00 PHST- 2019/09/02 00:00 [received] PHST- 2019/11/25 00:00 [revised] PHST- 2019/11/28 00:00 [accepted] PHST- 2019/12/04 06:00 [pubmed] PHST- 2020/04/09 06:00 [medline] PHST- 2019/12/04 06:00 [entrez] AID - S1050-4648(19)31110-6 [pii] AID - 10.1016/j.fsi.2019.11.067 [doi] PST - ppublish SO - Fish Shellfish Immunol. 2020 Jan;96:41-52. doi: 10.1016/j.fsi.2019.11.067. Epub 2019 Nov 30.