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 |
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| Main Authors: | , , , |
| Format: | Journal Article |
| Language: | English |
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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 |
| AuthorAffiliation_xml | – name: 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 |
| Author_xml | – sequence: 1 givenname: Xi surname: Gao fullname: Gao, Xi organization: School of Life Sciences/Hebei Basic Science Center for Biotic Interaction, Hebei University – sequence: 2 givenname: Mengmeng surname: Duan fullname: Duan, Mengmeng organization: School of Life Sciences/Hebei Basic Science Center for Biotic Interaction, Hebei University – sequence: 3 givenname: Yuying surname: Sun fullname: Sun, Yuying email: sunyuying125@hbu.edu.cn organization: School of Life Sciences/Hebei Basic Science Center for Biotic Interaction, Hebei University, Institute of Life Science and Green Development, Hebei University – sequence: 4 givenname: Jiquan surname: Zhang fullname: Zhang, Jiquan email: zhangjiquan@hbu.edu.cn organization: School of Life Sciences/Hebei Basic Science Center for Biotic Interaction, Hebei University |
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| Copyright | Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2024 Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2024. Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
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| Keywords | molting cycle RNAi calcium-associated cuticular protein calcium-binding ability Neocaridina denticulata sinensis |
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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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| Title | Molecular cloning and functional analysis of two calcium-associated cuticular protein genes in Neocaridina denticulata sinensis |
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