Fishery College, Guangdong Ocean University, Zhanjiang, China.
Fishery College, Guangdong Ocean University, Zhanjiang, China; Guangdong Technology Research Center for Pearl Aquaculture and Process, Guangdong Ocean University, Zhanjiang, China.
Comp Biochem Physiol B Biochem Mol Biol. 2020 Feb;240:110374. doi: 10.1016/j.cbpb.2019.110374. Epub 2019 Nov 14.
Chitin participates in shell formation as the main component of an organic framework. Chitin-binding protein contains domains that can bind to chitin specifically. In this study, a novel chitin-binding protein from Pinctada fucata martensii (PmCBP) with poly (chitin-binding domain) was cloned, which contains a 5'-untranslated region (UTR) of 114 bp and 3'UTR of 116 bp, and encodes a putative protein of 2044 amino acids. The predicted PmCBP protein was structurally typical of the CBP family with 20 ChtBD2 domains. Phylogenetic and linear relation analyses showed that the ChtBD2 domain has a highly conserved structure among the three species of P. f. martensii, Crassostrea gigas, and Mizuhopecten yessoensis. qRT-PCR and in-situ hybridization analysis revealed that PmCBP was most abundant in the mantle pallium whose expression level was significantly correlated with the growth traits. After RNAi, PmCBP expression was significantly inhibited in the mantle pallium (P < 0.05) and the microstructure of nacreous layers showed a disordered growth in the experiment group. These results indicated that PmCBP may be involved in nacreous layer formation through participation in the process of binding chitin in pearl oyster P. f. martensii.
几丁质作为有机框架的主要成分参与贝壳形成。几丁质结合蛋白含有可以特异性结合几丁质的结构域。在这项研究中,从马氏珠母贝(Pinctada fucata martensii)克隆到一种新型的几丁质结合蛋白 PmCBP,其包含 114bp 的 5'-非翻译区(UTR)和 116bp 的 3'-UTR,编码一个由 2044 个氨基酸组成的假定蛋白。预测的 PmCBP 蛋白在结构上典型地属于 CBP 家族,具有 20 个 ChtBD2 结构域。系统发育和线性关系分析表明,ChtBD2 结构域在马氏珠母贝、中国蛤蜊和虾夷扇贝这三个物种中具有高度保守的结构。qRT-PCR 和原位杂交分析显示,PmCBP 在套膜边缘组织中含量最丰富,其表达水平与生长性状显著相关。在 RNAi 后,PmCBP 在套膜边缘组织中的表达显著受到抑制(P<0.05),实验组珍珠层的微观结构显示出无序的生长。这些结果表明,PmCBP 可能通过参与珠母贝中几丁质的结合过程而参与珍珠层的形成。