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Identification and molecular characterization of a novel anti-lipopolysaccharide factor (ALF) from red swamp crayfish, Procambarus clarkii

Author:
Zhu, Juan-Juan, Ye, Zhi-Ze, Li, Chang-Sheng, Kausar, Saima, Abbas, Muhammad Nadeem, Xiang, Guang-Heng, Qian, Xing-Yi, Dai, Li-Shang
Source:
International journal of biological macromolecules 2019 v.132 pp. 43-50
ISSN:
0141-8130
Subject:
Escherichia coli, Procambarus clarkii, RNA interference, antibacterial properties, antiviral properties, cysteine, gene expression, genes, hepatopancreas, innate immunity, messenger RNA, nucleotide sequences, pathogens, peptidoglycans, polyclonal antibodies, polyinosinic-polycytidylic acid, quantitative polymerase chain reaction, rabbits, recombinant proteins, reverse transcriptase polymerase chain reaction, signal peptide, tissues
Abstract:
Anti-lipopolysaccharide factors are a group of small proteins with broad spectrum antiviral property and antibacterial activity. Herein, we obtained the genomic sequence of the Procambarus clarkii anti-lipopolysaccharide factor (PcALF) gene by using polymerase chain reaction to investigate its expression pattern in various tissues and in the immune tissues (Hepatopancreas) following exposure to pathogens. The deduced protein of PcALF was conserved; it displayed the signal peptides and putative lipo-polysaccharide binding domain, particularly the two conserved cysteine amino acid residues at both ends of the domain. The recombinant protein of PcALF was successfully expressed in Escherichia coli and rabbit anti-PcALF polyclonal antibodies were prepared. The qRT-PCR analysis showed unequal distribution of PcALF transcript in the examined tissues, however the transcript level was greatest in hepatopancreas. The challenge with peptidoglycan (PGN), lipo-polysaccharide (LPS) and Poly I:C significantly enhanced expression level of PcALF in hepatopancreas when compared with the PBS control. RNA interference of PcALF affected the mRNA expression levels of immune-related genes. Taken together, our data suggested that PcALF is an inducible protein and could play a key biological role in the innate immune defense of P. clarkii.
Agid:
6358326