Molecular Cloning and Expression of AjCaspase3-p Gene in Sea Cucumber Apostichopus japonicus
WANG Luyao1,2, ZHANG Chanchan2, ZHANG Xiaojing2, HAI Hangyu2, MA Deyou1,2
1. College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China; 2. Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture and Rural Affairs, Dalian 116023, China
Abstract：In order to explore the biological function of Caspase-3 gene as the crucial executioner protein in metamorphosis of sea cucumbers Apostichopus japonicus, a full-length cDNA of caspase-3 gene was cloned by cDNA end rapid amplification (RACE) technology, and the characteristics were analyzed. The expression levels of the gene were examined in various tissues and different development stages of thelarvae in the sea cucumber by RT-qPCR. Meanwhile, its spatial distribution was localized during metamorphosis process by the whole-amount in situ hybridization method. The results showed that the cloned Caspase-3 cDNA had full length of 2007 bp encoding 508 amino acids. Blastp comparison revealed that the gene had the most similarity with the putative caspase-3 gene (PIK46501.1) fromthe sea cucumbergenome designed as AjCaspase3-p. Bioinformatics analysis indicated that AjCaspase3-p contained the CASc domain unique to the Caspase family and a conserved QACRG cysteine active site. Phylogenetic analysis showed that AjCaspase3-p had high homology with Caspase-3 of other species belonged to marine invertebrates. There was a constitutive expression in detected tissues, with the maximal expression level in coelomocytes (P<0.05), suggesting that AjCaspase3-p might be involved in host immune defense. The AjCaspase3-p mRNA expression level was also increased at late auricularia stage with peak at juvenile stage. Furthermore, spatial localization results showed that the expression positive reactivity of this gene gradually became obvious during metamorphosis, and was strongly expressed in tentacles and intestines at juvenils stage, indicating that the AjCaspase3-p might maintain homeostasis to ensure metamorphosis of the sea cucumber larvae.
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2025, Vol. 44 
