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cDNA cloning, expression and antibacterial activity of lysozyme c in the blue shrimp (Litopenaeus stylirostris)

Progress in Natural Science:Materials International 2009年第7期19卷 837-844页

作者： Weijun Mai a,b, chaoqun Hu a, a Applied Marine Biology Laboratory, South china Sea Institute of Oceanology, Science Academy of china, Guangzhou 510301, china b Graduate University of chinese Academy of Sciences, Beijing 100049, china Applied Marine Biology Laboratory South China Sea Institute of Oceanology Science Academy of China Guangzhou 510301 China Graduate University of chinese Academy of Sciences Beijing 100049 China

The gene coding for lysozyme in blue shrimp (Litopenaeus stylirostris) was cloned, sequenced and expressed in pET-32a vector. The deduced amino acid sequence of F. merguiensi lysozyme showed 37–93% similarity with the mouse, human, chicken, and tiger prawn counterparts. The lysozyme was purified to homogeneity and observed as a band of approximately 15 kDa in 15% SDS–PAGE. Semiquantitative RT-PcR analysis demonstrated that mRNA transcripts of lysozyme could be mainly detected in the tissues of haemocytes, gill, gonad and the lymphoid organ of unchallenged shrimps, whereas the expression of lysozyme transcripts was increased in all the tested tissues after the heat-killed Vibrio alginolyticus challenge. The temporal expression of lysozyme mRNA in haemolymph challenged by Micrococcus luteus and V. alginolyticus was both up-regulated and reached the maximum level at 8 and 16 h post-stimulation, respectively, and then dropped back to the original level. Bacteriolytic activity of the lysozyme against different bacterial cultures was determined by the solid phase and turbidimetric assays. The results demonstrated that the lysozyme we obtained was not only against Grampositive and Gram-negative bacteria but also against shrimp pathogens V. alginolyticus and V. parahemolyticus. In addition, the study of the inhibition mechanism revealed that the antibacterial activity of the lysozyme was a result of the bactericidal effect.

关键词： lysozyme c Litopenaeus stylirostris PcR-cloning Expression Antibacterial activity

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Active transcription of MHc class I and lysozyme c in response of crucian carp（carassius auratus gibelio） to cyprinid herpesvirus 2 infection: implication for their basic role in antiviral innate immunity

Active transcription of MHC class I and Lysozyme C in respon...

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2014年中国水产学会学术年会

作者： Patarida Podok Lijuan Xu Dan Xu Liqun Lu Key Laboratory of Aquatic Genetic Resources of the Ministry of Agriculture Shanghai Ocean University

MHc class I（MHc I） and lysozyme c（LYZ c） are play important roles associated with disease resistant in fish species and believed to play in antiviral innate immunity. Through suppression subtractive hybridization（SSH） analysis, we previously identified MHc I and LYZ c as the two genes that were expressed in both of moribund and survivor SSH libraries after cyprinid herpesvirus 2（cy HV-2） infections. Here, we have further characterized two genes and their response to viral challenge The open reading frames（ORF） of MHc I and LYZ c were cloned from crucian carp（carassius auratus gibelio）. The full-length c DNAs of the two genes contained an ORF of 1,038 and 438 bp, encode a polypeptide of 345 and 145 amino acids, respectively. Phylogenetic analysis indicated that MHc I and LYZ c were most closely related to common carp. We found that two genes were expressed at low levels in all of the tissues examined in healthy crucian carp. The m RNA expression of MHc I and LYZ c in crucian carp kidney after cy HV-2 infection by using real-time RT PcR analysis at different time points, during challenge experiments, the peak of MHc I was observed at 72 hpi.（5.55-fold, P＜0.01）. While, 48 hpi（3.87-fold, P＜0.01） was the highest expression time points for LYZ c. For the fish died and survived from viral infection, expression levels of MHc I were 3.17 fold and 3.05 fold（P＜0.01）, respectively. Same as in the expression levels of LYZ c, were significantly upregulated in both died and survived fish（2.32 fold and 2.56 fold（P＜0.05）, respectively）. These data suggested MHc I and LYZ c were involved in cellular response genes to cy HV-2 but not play a key role in resistance.

关键词： crucian carp carassius auratus gibelio innate immunity MHc class I lysozyme c

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