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

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...

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Published in:Journal of oceanology and limnology Vol. 42; no. 5; pp. 1659 - 1672
Main Authors: Gao, Xi, Duan, Mengmeng, Sun, Yuying, Zhang, Jiquan
Format: Journal Article
Language:English
Published: Heidelberg Science Press 01.09.2024
Springer Nature B.V
Institute of Life Science and Green Development,Hebei University,Baoding 071002,China
School of Life Sciences/Hebei Basic Science Center for Biotic Interaction,Hebei University,Baoding 071002,China%School of Life Sciences/Hebei Basic Science Center for Biotic Interaction,Hebei University,Baoding 071002,China
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ISSN:2096-5508, 2523-3521
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Abstract 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 NdBcR -mediated 20E 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.
AbstractList 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 D2–4 stage, NdCAP-2 expressed in D2–4 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 NdBcR-mediated 20E 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.
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 D2-4 stage,NdCAP-2 expressed inD24and 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 20E 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.
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 D₂–₄ stage, NdCAP-2 expressed in D₂–₄ 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 NdBcR-mediated 20E 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.
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 NdBcR -mediated 20E 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.
Author Zhang, Jiquan
Gao, Xi
Duan, Mengmeng
Sun, Yuying
AuthorAffiliation School of Life Sciences/Hebei Basic Science Center for Biotic Interaction,Hebei University,Baoding 071002,China%School of Life Sciences/Hebei Basic Science Center for Biotic Interaction,Hebei University,Baoding 071002,China;Institute of Life Science and Green Development,Hebei University,Baoding 071002,China
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Keywords molting cycle
RNAi
calcium-associated cuticular protein
calcium-binding ability
Neocaridina denticulata sinensis
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Institute of Life Science and Green Development,Hebei University,Baoding 071002,China
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Snippet The body surface of crustaceans is covered with a sturdy shell. The growth and development of crustaceans are realized through molting. Neocaridina denticulata...
The body surface of crustaceans is covered with a sturdy shell. The growth and development of crustaceans are realized through molting. Neocaridina denticulata...
The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata...
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SubjectTerms Animal cuticle
Aquatic crustaceans
Binding
Calcification
Calcium
Calcium carbonate
Calcium carbonates
Calcium ions
Carbonates
Cephalothorax
Chitin
Cloning
Crustacea
Crustaceans
Earth and Environmental Science
Earth Sciences
Epicuticle
Functional analysis
Genes
growth and development
Molecular docking
Molting
Moulting
Neocaridina denticulata
Nucleotide sequence
Oceanography
Proteins
Recombinants
Research Paper
reverse transcriptase polymerase chain reaction
Reverse transcription
Surface defects
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Title Molecular cloning and functional analysis of two calcium-associated cuticular protein genes in Neocaridina denticulata sinensis
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