XIONG Panpan, ZHANG Hua, HE Maoxian. Identification of IGF2BP1 and its effect on the expression of mineralization-related genes in Pinctada fucata martensii[J]. Journal of fisheries of china, 2022, 46(12): 2265-2273. DOI: 10.11964/jfc.20210412758
Citation: XIONG Panpan, ZHANG Hua, HE Maoxian. Identification of IGF2BP1 and its effect on the expression of mineralization-related genes in Pinctada fucata martensii[J]. Journal of fisheries of china, 2022, 46(12): 2265-2273. DOI: 10.11964/jfc.20210412758

Identification of IGF2BP1 and its effect on the expression of mineralization-related genes in Pinctada fucata martensii

  • Insulin-like growth factor 2 binding proteins (IGF2BPs) are the important RNA metabolism regulators involved in many different biological processes. In order to study whether IGF2BP1 plays a role in biomineralization of pearl oysters, this study identified a Pinctada fucata martensii IGF2BP1 gene, named PfIGF2BP1-1. The cDNA length of the gene is 7 348 bp, with an ORF of 1 818 bp and encoding 605 amino acids. Multiple sequence alignments and phylogenetic analysis showed that PfIGF2BP1-1 has two RRM domains and four KH domains, which are homologous to IGF2BPs from other species. Real-time fluorescent quantitative PCR results showed that PfIGF2BP1-1 was expressed in eight tissues of P. fucata martensii, with the highest expression level in adductor muscle tissue, followed by foot and mantle tissue. The pET-32a prokaryotic expression vector was used to construct a recombinant plasmid containing the mature polypeptide region of PfIGF2BP1-1 and the protein was successfully expressed. The optimal conditions for inducing protein expression were that the bacteria were cultured at 37 ℃ for 8 hours with 0.5 mmol/L IPTG . The PfIGF2BP1-1 protein can stimulate the coat membrane primary cells of P. fucata martensii and significantly increase the expression level of cell matrix protein genes Accbp, MSI7, and Nacrein in primary mantle cells. The results indicated that the PfIGF2BP1-1 protein is involved in the biomineralization process of P. fucata martensii through inducing the expression of shell mineralization-related genes.
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