Cloning and functional analysis of the molting gene CYP302A1 of Daphnia sinensis.
Cloning and functional analysis of the molting gene CYP302A1 of Daphnia sinensis.
复制标题
中华溞蜕皮基因CYP302A1的克隆及功能分析
DOI：
10.1186/s12983-023-00483-2
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发表时间：
2023-01-12
影响因子：
2.8
通讯作者：
Deng, Daogui
中科院分区：
文献类型：
--
作者：
Qi, Huiying;Cao, Huijuan;Zhao, Yajie;Cao, Yaqin;Jin, Qide;Wang, Yeping;Zhang, Kun;Deng, Daogui
背景
蜕皮是节肢动物生长发育过程中的重要生理过程，主要受保幼激素和蜕皮激素的调控。CYP 302 A1是昆虫蜕皮激素合成的关键酶，但在枝角类中尚未发现。
结果
本文克隆了中华蚤CYP 302 A1基因，并对其功能进行了分析。CYP 302 A1基因在D.结果表明，该基因全长5926 bp，开放阅读框（ORF）为1596 bp，编码531个氨基酸，分子量为60.82 kDa，等电点为9.29。氨基酸序列分析表明，该蛋白具有5个细胞色素P450家族特有的保守区（helix-C、helix-K、helix-I、PERF和血红素结合区）。在dsRNA介导的实验中，在5%大肠杆菌浓度下，CYP 302 A1基因的表达水平显著降低（敲低56.22%）。此外，下游的EcR、USP和HR 3基因的表达水平也显著降低，而FTZ-f1基因的表达水平显著升高。在5%E. coli处理后，D.中华绒螯蟹胚胎发育时间延长，孵化囊内胚胎发育出现异常或解体。整体原位杂交结果表明，CYP 302 A1基因在D. RNA干扰前中华绒螯蟹有6个表达位点，分别位于第一触角神经节、卵巢、盲肠、嗅毛、胸肢和尾棘。而D. RNA干扰后，中华绒螯蟹第一触角神经节中的RNA缺失，卵巢中RNA的表达明显减弱。
结论
CYP 302 A1基因在D.中华绒螯蟹（D. sinensis）的蜕皮和繁殖受到影响。sinensis。
Background
Molting is an important physiological process in the growth and development of arthropoda, which is mainly regulated by juvenile hormone and ecdysone. CYP302A1 is a key enzyme which plays a critical role in the synthesis of ecdysone in insects, but it has not been identified in cladocera.
Results
The CYP302Al gene of Daphnia sinensis was cloned and its function was analyzed in this paper. The CYP302Al gene of D. sinensis was 5926 bp in full-length, with an open reading frame (ORF) of 1596 bp that encoded 531 amino acids (aa), a molecular weight of 60.82 kDa and an isoelectric point of 9.29. The amino acid sequence analysis revealed that there were five characteristic conserved regions of cytochrome P450 family (namely helix-C, helix-K, helix-I, PERF and heme-binding). In dsRNA mediated experiment, the expression level of CYP302A1 gene decreased significantly (knock-down of 56.22%) in the 5% Escherichia coli concentration treatment. In addition, the expression levels of EcR and USP and HR3 genes in the downstream were also significantly decreased, whereas that of FTZ-f1 gene increased significantly. In the 5% E. coli treatment, the molting time at maturity of D. sinensis prolonged, and the development of embryos in the incubation capsule appeared abnormal or disintegrated. The whole-mount in situ hybridization showed that the CYP302A1 gene of D. sinensis had six expression sites before RNA interference (RNAi), which located in the first antennal ganglion, ovary, cecae, olfactory hair, thoracic limb and tail spine. However, the expression signal of the CYP302A1 gene of D. sinensis disappeared in the first antennal ganglion and obviously attenuated in the ovary after RNAi.
Conclusion
The CYP302A1 gene played an important role in the ecdysone synthesis pathway of D. sinensis, and the knock-down of the gene affected the molting and reproduction of D. sinensis.
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影响因子：
3.5
作者：
Hiruta C;Toyota K;Miyakawa H;Ogino Y;Miyagawa S;Tatarazako N;Shaw JR;Iguchi T
通讯作者：
Iguchi T
影响因子：
4.1
作者：
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通讯作者：
Gilbert, LI
影响因子：
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作者：
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通讯作者：
Kataoka, H
影响因子：
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作者：
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通讯作者：
Zhao, Yunlong
影响因子：
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作者：
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通讯作者：
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