Journal of Tropical Oceanography ›› 2020, Vol. 39 ›› Issue (3): 66-75.doi: 10.11978/2019074CSTR: 32234.14.2019074
• Marine Biology • Previous Articles Next Articles
Cloning and expression analysis of aquaporin gene AQP4 cDNA from Pinctada fucata martensii
Xiaolan PAN1,2,3, Huiru LIU1,2,3, Meng XU1,2,3, Hanzhi XU1,2,3, Hua ZHANG1,4, Maoxian HE1,4(
)
- 1. CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
2. Guangdong Provincial Key Laboratory of Applied Marine Biology, Guangzhou 510301, China
3. University of Chinese Academy of Sciences, Beijing 100049, China
4. Institution of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences, Guangzhou 510301, China
-
Received:2019-08-19Revised:2019-11-28Online:2020-05-10Published:2020-05-19 -
Contact:Maoxian HE E-mail:hmx2@scsio.ac.cn -
Supported by:The Earmarked Fund for Modern Agro-industry Technology Research System(CARS-49);Marine Ecological Civilization Project(ISEE2018PY03);Science and Technology Planning Project of Guangdong Province, China(2017B0303014052)
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Xiaolan PAN, Huiru LIU, Meng XU, Hanzhi XU, Hua ZHANG, Maoxian HE. Cloning and expression analysis of aquaporin gene AQP4 cDNA from Pinctada fucata martensii[J].Journal of Tropical Oceanography, 2020, 39(3): 66-75.
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Tab. 1
Primers used in this study"
| 引物名称 | 引物序列(5′—3′) | 注释 |
|---|---|---|
| GSP5 | CGATAGCCCAAAGGCAAAAGATA | 5′RACE外引物 |
| NGSP5 | GAAGGTGGCAGAGGTACTCTTGAAGG | 5′RACE内引物 |
| GSP3 | GTTCGTCTTTGCTGCTCCTTCC | 3′RACE外引物 |
| NGSP3 | GACCTGAAGAAGGACGAAAACG | 3′RACE内引物 |
| Faqp | CTGTGATGGCGTCAACTGATG | PfAQP4定量引物 |
| Raqp | CGCAATCGTGAGAAGATGACC | |
| F18S | CGTTTCAACAAGACGCCAGTAG | 内参基因18S定量引物 |
| R18S | ACGAAAAAAAGGTTTGAGAGACG |
Tab. 1
Fig. 1
cDNA sequence and deduced amino acid sequence of PfAQP4. Lower case letters indicate 5'UTR and 3'UTR, upper case letters indicate ORF, shaded areas indicate SMART-predicted MIP domains, boxes indicate NPA sequences, underlined letters indicate six transmembrane regions, and * represents protein translation terminator. aataaa in bold represents polyadenylation tail signal"
Fig. 1
Fig. 2
Hydrophobic plots of PfAQP4. Numbers 1-6 represents six transmembrane regions, A-E represent five loops, and the windows size refers to the interval length used for contour calculation. When calculating the score of a given residue i, the amino acids in the selected length interval centered on residue i are considered"
Fig. 2
Fig. 3
Predicted 3D structure of PfAQP4 (a) and H. sAQP4 (b)"
Fig. 3
Tab. 2
Amino acid identity comparison of PfAQP4 with other known AQP4 homologues"
| 物种名 | 多序列比对用缩写名 | 登录号 | 一致性/% |
|---|---|---|---|
| 马氏珠母贝(Pinctada fucata martensii) | P.f | MN310685 | 100 |
| 斑马鱼(Danio rerio) | D.r | ACV73794.1 | 39.5 |
| 鱇浪白鱼(Anabarilius grahami) | A.g | ROL42342.1 | 37.9 |
| 斑点叉尾鮰(Ictalurus punctatus) | I.p | AHH42521.1 | 40.9 |
| 暗纹东方鲀(Takifugu obscurus) | T.o | BAL44699.1 | 40.3 |
| 小鼠(Mus musculus) | M.m | AAC53155.1 | 40.4 |
| 大鼠(Rattus norvegicus) | R.n | 2D57 | 41.1 |
| 智人(Homo sapiens) | H.s | 3GD8 | 42.2 |
| 非洲爪蟾(Xenopus tropicalis) | X.t | NP_001304774.1 | 39.9 |
| 长牡蛎(Crassostrea gigas) | C.g | EKC31493.1 | 53.4 |
| 虾夷扇贝(Mizuhopecten yessoensis) | M.y | OWF48037.1 | 52.2 |
| 停乳链球菌(Streptococcus dysgalactiae) | SQB67194.1 | 23.4 | |
| 犬链球菌(Streptococcus canis) | VEE24698.1 | 23.5 |
Tab. 2
Fig. 4
Multi-sequence alignment of PfAQP4 with other species AQP4. α1-α6 represents six transmembrane regions, the box represents the NPA motif, the white letter with black background indicates the column are conserved (≥50%), the gray shading indicates the columns are similar, and the black letter with white background indicates that the residues are not conserved"
Fig. 4
Fig. 5
NJ phylogenetic analysis of PfAQP4 with AQP4 of other species"
Fig. 5
Fig. 6
Relative expressions of PfAQP4 in different tissues. Different letters indicate significant differences between groups (P<0.01)"
Fig. 6
Fig. 7
Relative expressions of PfAQP4 at different developmental stages"
Fig. 7
Fig. 8
Relative expressions of PfAQP4 under different salinity stress. ** indicates that the same salinity group is significantly different at different times (P<0.01)"
Fig. 8
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