Journal of Dalian Ocean University 2023, Vol. 38 Issue (6): 947-955 DOI: 10.16535/j.cnki.dlhyxb.2023-111 |
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Expression and immune function characteristics of peptidoglycan recognition protein gene in disk abalone (Haliotis discus hannai) |
QIAO Kun,ZHENG Chunhua,CHEN Bei,SU Yongchang,HAO Hua,XU Min,CAI Shuilin,LIU Zhiyu*
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1.Key Laboratory of Cultivation and High-Value Utilization of Marine Organisms in Fujian Province,Fisheries Research Institute of Fujian,Xiamen 361013,China;2.College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China;3.College of Ocean and Earth Sciences,Xiamen University,Xiamen 361102,China |
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Abstract In order to investigate the immune function of the peptidoglycan recognition protein HdhPGRP-SC2-like gene in disk abalone (Haliotis discus hannai), the full-length cDNA sequence of HdhPGRP-SC2-like gene was cloned by RACE technology, and tissue distribution of HdhPGRP-SC2-like gene was analyzed in the disk abalone by real-time fluorescent quantitative PCR(qPCR). The recombinant protein rHdhPGRP-SC2-like was obtained by prokaryotic expression, and its amidase activity and immune characteristics were verified. The results showed that the cDNA of the HdhPGRP-SC2-like gene was 938 bp in length, with an open reading frame of 780 bp, encoding 260 amino acids and containing the conserved PGRP and Ami_2 structural domains. HdhPGRP-SC2-like gene as a short type PGRP had high similarity with the closely related species PGRP of mollusks. qPCR analysis showed that HdhPGRP-SC2-like mRNA was expressed in all tissues of the disk abalone, with the maximal expression level in gill, a certain amount of expression level in mantle and digestive gland, and the minimal content in foot and hemocyte. The prokaryotic expression vector pET-28a(+)-HdhPGRP-SC2-like was further constructed, and the recombinant protein rHdhPGRP-SC2-like gene was obtained by induction of expression, isolation, and purification. The recombinant protein had peptidoglycan binding activity and showed amidase activity in the presence of Zn2+. The finding indicated that HdhPGRP-SC2-like gene binded bacterial peptidoglycan components and played an important role in the immune defence against invading bacteria.
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Published: 18 January 2024
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