Molecular cloning and functional analysis of two calcium associated cuticular protein genes in Neocaridina denticulata sinensis  

作　　者：Xi GAO Mengmeng DUAN Yuying SUN Jiquan ZHANG 

机构地区：[1]School of Life Sciences/Hebei Basic Science Center for Biotic Interaction,Hebei University,Baoding 071002,China [2]Institute of Life Science and Green Development,Hebei University,Baoding 071002,China

出　　处：《Journal of Oceanology and Limnology》2024年第5期1659-1672,共14页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(Nos.32172954,32373121);the Key Research and Development Project of Hebei Province(No.22323201D);the Science and Technology Project of Hebei Education Department(No.ZD 2022093);the Natural Science Foundation of Hebei Province(Nos.D2022201003,D2023201002);the Hangzhou Qianjiang Special Expert fund for Prof.Jiquan ZHANG。

摘　　要：The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata sinensis is a suitable candidate for crustacean scientific research.Two calcium-associated cuticular protein genes,named NdCAP-1 and NdCAP-2,were obtained from N.denticulata sinensis.Molecular docking simulated the binding effect of both proteins and calcium ions.Semi-quantitative reverse transcription PCR results show that NdCAP-1 is expressed in D_(2-4) stage,NdCAP-2 expressed in D_(2-4) and A-B stages,and both were significantly expressed in the cephalothorax cuticle and pereiopods.Then,it was revealed that NdCAP-1 and NdCAP-2 are regulated by NdEcR-mediated 20 E signaling pathways.Knockdown of NdCAP-1 and NdCAP-2 was observed to cause surface defects.The recombinant proteins(rNdCAP-1 and rNdCAP-2),obtained by prokaryotic expression,had calcium-binding and chitin-binding ability,inhibited formation of calcium carbonate precipitate.These results show that calcium-associated cuticular proteins play important roles in cuticle formation and calcification.

关 键 词：Neocaridina denticulata sinensis calcium-associated cuticular protein molting cycle RNAi calcium-binding ability 

分 类 号：S917.4[农业科学—水产科学]